Anti-cd47 antibodies and methods of use thereof

ABSTRACT

The invention relates to monoclonal and/or monovalent antibodies that bind CD47. The invention relates to monoclonal and/or monovalent antibodies that bind CD19. The invention also relates to novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule, where one of the binding sites is specific for CD47. The invention also relates to novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule, where one of the binding sites is specific for CD19.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/095,395, filed Dec. 3, 2013, which claims the benefit of U.S.Provisional Application No. 61/732,452, filed Dec. 3, 2012; of U.S.Provisional Application No. 61/816,788, filed Apr. 28, 2013; of U.S.Provisional Application No. 61/863,106, filed Aug. 7, 2013; of U.S.Provisional Application No. 61/881,523, filed Sep. 24, 2013; and of U.S.Provisional Application No. 61/898,710, filed Nov. 1, 2013; each ofwhich is incorporated herein by reference in its entirety.

INCORPORATION BY REFERENCE OF SEQUENCE LISTING

The contents of the text file named “NOVI_030_001US_SeqList_ST25.txt”,which was created on Jun. 26, 2014 and is 272 KB in size, are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

The invention relates to monoclonal and/or monovalent antibodies thatbind CD47. The invention relates to monoclonal and/or monovalentantibodies that bind CD19. The invention also relates to novelbispecific monoclonal antibodies carrying a different specificity foreach binding site of the immunoglobulin molecule, where one of thebinding sites is specific for CD47. The invention also relates to novelbispecific monoclonal antibodies carrying a different specificity foreach binding site of the immunoglobulin molecule, where one of thebinding sites is specific for CD19.

BACKGROUND OF THE INVENTION

CD47 or Integrin-Associated-Protein (IAP) is a ubiquitous 50 kDatransmembrane glycoprotein with multiple functions in cell-cellcommunication. It interacts with multiple ligands, such as integrins,SIRPα (Signal Regulatory Protein alpha), SIRPγ and thrombospondins(Oldenborg, P. A., CD47: A Cell Surface Glycoprotein Which RegulatesMultiple Functions of Hematopoietic Cells in Health and Disease, ISRNHematol. 2013; 2013:614619; Soto-Pantoja D R, et al., Therapeuticopportunities for targeting the ubiquitous cell surface receptor CD47(2012), Expert Opin Ther Targets. 2013 January; 17(1):89-103; Sick E, etal., CD47 Update: a multifaced actor in the tumor microenvironment ofpotential therapeutic interest, Br J Pharmacol. 2012 December;167(7):1415-30). In the context of the innate immune system, CD47functions as a marker of self, transmitting an inhibitory “don't killme” signal through binding to SIRPα expressed by myeloid cells, such asmacrophages, neutrophils, and dendritic cells. The role of widespreadexpression of CD47 in the physiological situation is therefore toprotect healthy cells against the elimination by the innate immunesystem (Oldenborg P A, et al., CD47-Signal Regulatory Protein α (Sirpα)Regulates Fcγ and Complement Receptor-Mediated Phagocytosis, J Exp Med.2001 Apr. 2; 193(7):855-62; Mattias Olsson, Role of theCD47/SIRPα-interaction in regulation of macrophage phagocytosis,Department of Integrative Medical Biology, Section for Histology andCellBiology, Umeå University, Umeå, Sweden, Thesis; Oldenborg P A., Roleof CD47 in erythroid cells and in autoimmunity, Leuk Lymphoma. 2004July; 45(7): 1319-27; Oldenborg P A, et al., Role of CD47 as a Marker ofSelf on Red Blood Cells., Science. 2000 Jun. 16; 288(5473):2051-4; BrownE J, Frazier W A., integrin-associated protein (CD47) and its ligands.,Trends Cell Biol. 2001 March; 11(3):130-5).

Tumor cells hijack this immunosuppressive mechanism by overexpressingCD47, which efficiently helps them to escape immune surveillance andkilling by innate immune cells. (Majeti R, Ch et al., CD47 is an adverseprognostic factor and therapeutic antibody target on human acute myeloidleukemia stem cells, Cell. 2009 Jul. 23; 138(2):286-99; S. Jaiswal etal., CD47 is upregulated on circulating hematopoietic stem cells andleukemia cells to avoid phagocytosis., Cell. 2009 Jul. 23;138(2):271-85). CD47 expression is upregulated in most human cancers(e.g., NHL, AML, breast, colon, glioblastoma, glioma, ovarian, bladderand prostate cancers) and increased levels of CD47 expression clearlycorrelate with aggressive disease and poor survival. (Majeti R, et al.,Cell. 2009 Jul. 23; 138(2):286-99; S. Jaiswal et al., Cell. 2009 Jul.23; 138(2):271-85; Willingham S B, et al., The CD47-signal regulatoryprotein alpha (SIRPα) interaction is a therapeutic target for humansolid tumors, Proc Natl Acad Sci USA. 2012 Apr. 24; 109(17):6662-7; ChaoM P, et al., Therapeutic antibody targeting of CD47 eliminates humanacute lymphoblastic leukemia., Cancer Res. 2011 Feb. 15; 71(4):1374-84).

The widespread expression of CD47 in healthy tissues brings the questionof treatment safety and efficacy: First, targeting CD47 with aneutralizing monoclonal antibody (Mab) could affect healthy cells,resulting in severe toxicities as shown in preclinical studies with miceand cynomolgus monkeys (Willingham S B, et al., Proc Natl Acad Sci USA.2012 Apr. 24; 109(17):6662-7; Weiskopf K, et al., Engineered SIRPαVariants as Immunotherapeutic Adjuvants to Anticancer Antibodies,Science. 2013 Jul. 5; 341(6141):88-91). Second, even if severetoxicities could be avoided or mitigated by using alternative formats(Weiskopf K, et al., Science. 2013 Jul. 5; 341(6141):88-91), broadexpression of CD47 could still cause a rapid elimination of CD47-bindingmolecules through target-mediated drug disposition resulting in poorpharmacokinetics and decreased efficacy.

Accordingly, there exists a need for antibodies and therapeutics thatenable targeting of CD47 and overcome these obstacles.

SUMMARY OF THE INVENTION

The invention provides monoclonal antibodies that bind CD47. Theseantibodies are collectively referred to herein as anti-CD47 monoclonalantibodies or anti-CD47 mAbs. Preferably, the monoclonal antibodies arespecific for at least human CD47. In some embodiments, the monoclonalantibodies that recognize human CD47 are also cross-reactive for atleast one other non-human CD47 protein, such as, by way of non-limitingexample, non-human primate CD47, e.g., cynomolgus monkey CD47, and/orrodent CD47. In some embodiments, these anti-CD47 monoclonal antibodiesinhibit the interaction between CD47 and signal-regulatory protein alpha(SIRPα). In some embodiments, these anti-CD47 monoclonal antibodiesinhibit the interaction between human CD47 and human SIRPα. Theinvention also include antibodies that bind to the same epitope as ananti-CD47 monoclonal antibody disclosed herein and inhibits theinteraction between CD47 and SIRPα, e.g., between human CD47 and humanSIRPα.

The invention also provides monovalent antibodies and/or bispecificantibodies that include at least a first arm that is specific for CD47.Preferably, the monovalent antibodies and/or bispecific antibodies arespecific for at least human CD47. In some embodiments, the monovalentantibodies and/or bispecific antibodies that recognize human CD47 arealso cross-reactive for at least one other non-human CD47 protein, suchas, by way of non-limiting example, non-human primate CD47, e.g.,cynomolgus monkey CD47, and/or rodent CD47. In some embodiments, theseanti-CD47 monovalent antibodies and/or anti-CD47 bispecific antibodiesinhibit the interaction between CD47 and signal-regulatory protein alpha(SIRPα). In some embodiments, these anti-CD47 monovalent antibodiesand/or anti-CD47 bispecific antibodies inhibit the interaction betweenhuman CD47 and human SIRPα. The invention also include antibodies thatbind to the same epitope as an anti-CD47 monovalent and/or an anti-CD47bispecific antibody disclosed herein and inhibits the interactionbetween CD47 and SIRPα, e.g., between human CD47 and human SIRPα.

The invention provides bispecific antibodies that recognize CD47 and asecond target. The invention allows for the identification, productionand purification of bispecific antibodies that are undistinguishable insequence from standard antibodies and where one of the binding sites isspecific for CD47 and the second binding site is specific for anothertarget, for example a tumor-associated antigen (TAA). In someembodiments, the TAA is an antigen that is expressed on the cell surfaceof a cancer cell. In some embodiments, the cancer cell is selected froma lung cancer cell, a bronchial cancer cell, a prostate cancer cell, abreast cancer cell, a colorectal cancer cell, a pancreatic cancer cell,an ovarian, a leukemia cancer cell, a lymphoma cancer cell, anesophageal cancer cell, a liver cancer cell, a urinary and/or bladdercancer cell, a renal cancer cell, an oral cavity cancer cell, apharyngeal cancer cell, a uterine cancer cell, and/or a melanoma cancercell.

In some embodiments, suitable TAA, by way of non-limiting example,include CD20, HER2, HER3, EGFR, IGF1R, c-Met, PDGFR1, CD40, CD40L, CD30,CS1, CD70, glypican, mesothelin, PSMA, PSCA, MUC1, CA125, CEA, FRA,EpCAM, DR5, HGFR1, and/or 5T4.

CD47 (Cluster of Differentiation 47) functions as a “don't eat me”signal for phagocytic cells and is known to be over-expressed by manytumors (immune escape). CD47 interacts with SIRPα, which is expressed onphagocytic cells. CD47 down-regulates phagocytic activity. CD47 inhibitsdendritic cell (DC) maturation and activation. CD47 has also beenimplicated in processes such as, for example, apoptosis, survival,proliferation, adhesion, migration, and regulation of angiogenesis,blood pressure, tissue perfusion, and/or platelet homeostasis.

CD47 has also been implicated in cancer. For example, CD47 isoverexpressed in various hematological and solid malignancies. CD47 is adocumented cancer stem cell/tumor initiating cell marker. It is thoughtthat CD47 overexpression may help tumor cells to escape immunesurveillance and killing by innate immune cells. High levels of CD47 arealso associated with poor clinical outcome in cancers such as, forexample, leukemias, lymphomas, breast cancer, colon cancer, ovariancancer, bladder cancer, prostate cancer, and/or glioma. Thus, targetingCD47 would be useful in treating, delaying the progression of, orotherwise ameliorating a symptom of cancer.

As CD47 is ubiquitously expressed, it is a difficult target for amonoclonal antibody (mAb). Nevertheless, the antibodies that arespecific for CD47 described herein are useful as monospecific antibodiesand can be used for therapeutic intervention or as a research ordiagnostic reagent. Monospecific antibodies of the invention that bindCD47, as well as fragments of these monospecific antibodies that areimmunologically active and still bind CD47, include the exemplaryantibodies described herein, e.g., the 5A3 antibody, the 5A3M4 antibody,the 5A3M3 antibody, the 5A3M5 antibody, the KE8 antibody, the KE8-P6H5antibody (also referred to herein as KE8H5), the KE8-P3B2 antibody (alsoreferred to herein as KE8B2), the KE8-P2A2 antibody (also referred toherein as KE8A25), the KE8F2 antibody, the KE8G2 antibody, the KE84G9antibody, the KE81G9 antibody, the KE81A3 antibody, the KE8E8 antibody,the KE8G6 antibody, the KE8H3 antibody, the KE8C7 antibody, the KE8A4antibody, the KE8A8 antibody, the KE8G11 antibody, the KE8B7 antibody,the KE8F1 antibody, the KE8C4 antibody, the KE8A3 antibody, the KE86G9antibody, the KE8H6 antibody, the KA3 antibody, the KA3-P5G2 antibody(also referred to herein as KA3G2), the KA3-P1A3 antibody (also referredto herein as KA3A3), the KA3-P5C5 antibody (also referred to herein asKA3C5), the KA3H8 antibody, the KA3B2 antibody, the KA3A2 antibody, theKA3D3 antibody, the KA3H3 antibody, the KC4 antibody, theKC4-P1G11KC4-P4C11 antibody, the KC4-P6B1KC4-P4F4 antibody, and theKC4-P2E2 antibody (also referred to herein as KC4E2), the KC4 antibody,the KC4F4 antibody, the KC4A1 antibody, the KC4C11 antibody, the KC4E10antibody, the KC4B1 antibody, the KC4C3 antibody, the KC4A4 antibody,the KC4G11 antibody, the KC4G9 antibody and fragments thereof.

The antibodies of the invention that bind CD47 and fragments thereofserve to modulate, block, inhibit, reduce, antagonize, neutralize orotherwise interfere with the functional activity of CD47. Functionalactivities of CD47 include, by way of non-limiting example, interactionwith SIRPα. The antibodies are considered to completely modulate, block,inhibit, reduce, antagonize, neutralize or otherwise interfere with theCD47-SIRPα interaction when the level of CD47-SIRPα interaction in thepresence of the antibody is decreased by at least 95%, e.g., by 96%,97%, 98%, 99% or 100% as compared to the level of CD47-SIRPα interactionin the absence of binding with an antibody described herein. Theantibodies are considered to partially modulate, block, inhibit, reduce,antagonize, neutralize or otherwise interfere with the CD47-SIRPαinteraction when the level of CD47-SIRPα interaction in the presence ofthe antibody is decreased by less than 95%, e.g., 10%, 20%, 25%, 30%,40%, 50%, 60%, 75%, 80%, 85% or 90% as compared to the level ofCD47-SIRPα interaction in the absence of binding with an antibodydescribed herein.

The invention also provides bispecific antibodies in which at least onebinding site is specific for CD47. The bispecific antibodies of theinvention target CD47 and a second antigen, e.g., a tumor-associatedantigen (TAA). In some embodiments, the bispecific antibody includes afunctional Fc portion. The TAA-binding arm of the bispecific antibodytargets the CD47 arm to the tumor cell or cancer stem cell. The CD47 armblocks, inhibits or otherwise reduces the interaction between CD47 andSIRPα, thereby conveying an “eat me” signal to the phagocyte. In someembodiments, the TAA-binding arm of the bispecific antibody includes ananti-CD19 antibody sequence or antigen-binding fragment thereof.

In some embodiments, the bispecific antibody exhibits a “balanced”affinity for each of the two targets. In other embodiments, thebispecific antibody exhibits an “unbalanced” affinity for each of thetwo targets. For example, in an anti-CD47/CD19 bispecific antibody, theaffinity of the anti-CD19 arm is increased. For example, in ananti-CD47/CD19 bispecific antibody, the affinity of the anti-CD47 arm isdecreased. For example, in an anti-CD47/CD19 bispecific antibody, theaffinity of the anti-CD19 arm is increased and the affinity of theanti-CD47 arm is decreased. These unbalanced affinity bispecificantibodies are useful, for example, to improve selectivity for a targetcell or group of target cells.

In some embodiments, the affinity of the anti-CD19 arm is increased byat least 100 fold following affinity maturation. In some embodiments,the affinity of the anti-CD47 arm is decreased by at least 2 foldfollowing affinity dematuration. For example, in some embodiments, theanti-CD47 arm exhibits an affinity for CD47 that is between about 2 foldand 100 fold lower following affinity dematuration.

The bispecific antibodies of the invention that include at least oneanti-CD47 arm serve to modulate, block, inhibit, reduce, antagonize,neutralize or otherwise interfere with the functional activity of CD47.Functional activities of CD47 include, by way of non-limiting example,interaction with SIRPα. The bispecific antibodies are considered tocompletely modulate, block, inhibit, reduce, antagonize, neutralize orotherwise interfere with the CD47-SIRPα interaction when the level ofCD47-SIRPα interaction in the presence of the bispecific antibody isdecreased by at least 95%, e.g., by 96%, 97%, 98%, 99% or 100% ascompared to the level of CD47-SIRPα interaction in the absence ofbinding with a bispecific antibody described herein. The bispecificantibodies are considered to partially modulate, block, inhibit, reduce,antagonize, neutralize or otherwise interfere with the CD47-SIRPαinteraction when the level of CD47-SIRPα interaction in the presence ofthe bispecific antibody is decreased by less than 95%, e.g., 10%, 20%,25%, 30%, 40%, 50%, 60%, 75%, 80%, 85% or 90% as compared to the levelof CD47-SIRPα interaction in the absence of binding with a bispecificantibody described herein.

The anti-CD47 arms of the bispecific antibodies of the invention areuseful with a number of arms that bind other antigens, e.g., TAAs.Exemplary anti-CD47 arms, anti-CD47 monovalent antibodies and/orbispecific antibodies of the invention include the antibodies referredto herein as the 5A3 antibody, the 5A3M4 antibody, the 5A3M3 antibody,the 5A3M5 antibody, the KE8 antibody, the KE8-P6H5 antibody (alsoreferred to herein as KE8H5), the KE8-P3B2 antibody (also referred toherein as KE8B2), the KE8-P2A2 antibody (also referred to herein asKE8A25), the KE8F2 antibody, the KE8G2 antibody, the KE84G9 antibody,the KE81G9 antibody, the KE81A3 antibody, the KE8E8 antibody, the KE8G6antibody, the KE8H3 antibody, the KE8C7 antibody, the KE8A4 antibody,the KE8A8 antibody, the KE8G11 antibody, the KE8B7 antibody, the KE8F1antibody, the KE8C4 antibody, the KE8A3 antibody, the KE86G9 antibody,the KE8H6 antibody, the KA3 antibody, the KA3-P5G2 antibody (alsoreferred to herein as KA3G2), the KA3-P1A3 antibody (also referred toherein as KA3A3), the KA3-P5C5 antibody (also referred to herein asKA3C5), the KA3H8 antibody, the KA3B2 antibody, the KA3A2 antibody, theKA3D3 antibody, the KA3H3 antibody, the KC4 antibody, theKC4-P1G11KC4-P4C11 antibody, the KC4-P6B1KC4-P4F4 antibody, and theKC4-P2E2 antibody (also referred to herein as KC4E2), the KC4 antibody,the KC4F4 antibody, the KC4A1 antibody, the KC4C11 antibody, the KC4E10antibody, the KC4B1 antibody, the KC4C3 antibody, the KC4A4 antibody,the KC4G11 antibody, the KC4G9 antibody and fragments thereof. In someembodiments, the TAA-binding arm of the bispecific antibody includes ananti-CD19 antibody sequence or antigen-binding fragment thereof.

The invention provides isolated bispecific antibodies having a first armthat includes a first amino acid sequence that binds CD47 and a secondarm that includes a second amino acid sequence that does not bind CD47,wherein the bispecific antibody inhibits interaction between CD47 andsignal-regulatory protein alpha (SIRPα). In some embodiments, the secondamino acid sequence binds a tumor associated antigen (TAA). In someembodiments, the bispecific antibody inhibits interaction between humanCD47 and human SIRPα.

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα at a level that is at least ten timesmore potent than a corresponding level of inhibition of human CD47/humanSIRPα interaction exhibited by a monovalent anti-CD47 antibody thatincludes the first amino acid sequence that binds CD47 and a secondamino acid sequence that does not bind a human protein.

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα at a level that is at least 100 timesmore potent than a corresponding level of inhibition of human CD47/humanSIRPα interaction exhibited by a monovalent anti-CD47 antibody thatincludes the first amino acid sequence that binds CD47 and a secondamino acid sequence that does not bind a human protein.

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα at a level that is at least 1,000times more potent than a corresponding level of inhibition of humanCD47/human SIRPα interaction exhibited by a monovalent anti-CD47antibody that includes the first amino acid sequence that binds CD47 anda second amino acid sequence that does not bind a human protein.

In some embodiments, the bispecific antibody includes a first arm thatinhibits the interaction between human CD47 at the surface of cells andsoluble human SIRPα with an IC50 greater than 5 nM in the assaydescribed in Example 4 and in which the monovalent antibody 5A3M3 has anIC50 of approximately 13 nM.

In some embodiments, the bispecific antibody includes a first arm thatis recovered at more than 80% after incubation at 37° C. for 30 minutesin human whole blood at a concentration of 10 μg/ml as described inExample 15.

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα at a level that is at least tentimes, at least 100 times or at least 1,000 times more potent than acorresponding level of inhibition of human CD47/human SIRPα interactionexhibited by a monovalent anti-CD47 antibody that includes the firstamino acid sequence that binds CD47 and a second amino acid sequencethat does not bind a human protein, and includes a first arm thatinhibits the interaction between human CD47 at the surface of cells andsoluble human SIRPα with an IC50 greater than 5 nM in the assaydescribed in Example 4 and in which the monovalent antibody 5A3M3 has anIC50 of approximately 13 nM.

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα at a level that is at least tentimes, at least 100 times or at least 1,000 times more potent than acorresponding level of inhibition of human CD47/human SIRPα interactionexhibited by a monovalent anti-CD47 antibody that includes the firstamino acid sequence that binds CD47 and a second amino acid sequencethat does not bind a human protein, and includes a first arm that isrecovered at more than 80% after incubation at 37° C. for 30 minutes inhuman whole blood at a concentration of 10 μg/ml as described in Example15.

In some embodiments, the TAA is CD19. In some embodiments, the secondamino acid sequence does not bind a human protein.

In some embodiments, the first amino acid sequence includes a variableheavy chain complementarity determining region 1 (CDRH1) amino acidsequence of SEQ ID NO: 225, a variable heavy chain complementaritydetermining region 2 (CDRH2) amino acid sequence of SEQ ID NO: 226, avariable heavy chain complementarity determining region 3 (CDRH3) aminoacid sequence of SEQ ID NO: 227, a variable light chain complementaritydetermining region 1 (CDRL1) amino acid sequence selected from SEQ IDNO: 228-241 and 262-272, a variable light chain complementaritydetermining region 2 (CDRL2) amino acid sequence selected from 242-245and 273-280, and a variable light chain complementarity determiningregion 3 (CDRH3) amino acid sequence selected from 246-261 and 281.

In some embodiments, the first amino acid sequence includes a variableheavy chain amino acid sequence of SEQ ID NO: 114 and a variable lightchain amino acid sequence selected from SEQ ID NO: 116, 118, 120, 122,124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150,152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178,180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204 and 206.

In some embodiments, the bispecific antibody includes two copies of asingle heavy chain polypeptide and a first light chain and a secondlight chain, wherein the first and second light chains are different.

In some embodiments, at least a portion of the first light chain is ofthe Kappa type and at least a portion of the second light chain is ofthe Lambda type. In some embodiments, the first light chain includes atleast a Kappa constant region. In some embodiments, the first lightchain further includes a Kappa variable region. In some embodiments, thefirst light chain further includes a Lambda variable region. In someembodiments, the second light chain includes at least a Lambda constantregion. In some embodiments, the second light chain further includes aLambda variable region. In some embodiments, the second light chainfurther includes a Kappa variable region. In some embodiments, the firstlight chain includes a Kappa constant region and a Kappa variableregion, and wherein the second light chain includes a Lambda constantregion and a Lambda variable region.

In some embodiments, the constant and variable framework regionsequences are human.

The invention also provides bispecific antibodies and/or monovalentantibodies that include at least a first arm that inhibits theinteraction between human CD47 at the surface of cells and soluble humanSIRPα with an IC50 greater than 5 nM in the assay described in Example 4and in which the antibody 5A3M3 has an IC50 of approximately 13 nM.

The invention also provides bispecific antibodies and/or monovalentantibodies that include at least a first arm that is recovered at morethan 80% after incubation at 37° C. for 30 minutes in human whole bloodat a concentration of 10 μg/ml as described in Example 15. In someembodiments, the bispecific antibody and/or monovalent antibody inhibitsinteraction between CD47 and signal-regulatory protein alpha (SIRPα). Insome embodiments, the bispecific antibody and/or monovalent antibodyinhibits interaction between human CD47 and human SIRPα.

The invention also provides isolated bispecific antibodies having afirst arm that includes a first amino acid sequence that binds CD47 anda second arm that includes a second amino acid sequence that binds CD19,wherein the bispecific antibody inhibits interaction between CD47 andsignal-regulatory protein alpha (SIRPα).

In some embodiments, the bispecific antibody inhibits interactionbetween human CD47 and human SIRPα. In some embodiments, the bispecificantibody inhibits interaction between human CD47 and human SIRPα at alevel that is selected from the group consisting of at least ten timesmore potent, at least 100 times more potent and at least 1,000 timesmore potent than a corresponding level of inhibition of human CD47/humanSIRPα interaction exhibited by a monovalent anti-CD47 antibody thatincludes the first amino acid sequence that binds CD47 and a secondamino acid sequence that does not bind a human protein.

In some embodiments, the first amino acid sequence includes a variableheavy chain complementarity determining region 1 (CDRH1) amino acidsequence of SEQ ID NO: 225, a variable heavy chain complementaritydetermining region 2 (CDRH2) amino acid sequence of SEQ ID NO: 226, avariable heavy chain complementarity determining region 3 (CDRH3) aminoacid sequence of SEQ ID NO: 227, a variable light chain complementaritydetermining region 1 (CDRL1) amino acid sequence selected from SEQ IDNO: 228-241 and 262-272, a variable light chain complementaritydetermining region 2 (CDRL2) amino acid sequence selected from 242-245and 273-280, and a variable light chain complementarity determiningregion 3 (CDRH3) amino acid sequence selected from 246-261 and 281.

In some embodiments, the first amino acid sequence includes a variableheavy chain amino acid sequence of SEQ ID NO: 114 and a variable lightchain amino acid sequence selected from SEQ ID NO: 116, 118, 120, 122,124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150,152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178,180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204 and 206.

In some embodiments, the bispecific antibody includes two copies of asingle heavy chain polypeptide and a first light chain and a secondlight chain, wherein the first and second light chains are different.

In some embodiments, at least a portion of the first light chain is ofthe Kappa type and at least a portion of the second light chain is ofthe Lambda type. In some embodiments, the first light chain includes atleast a Kappa constant region. In some embodiments, the first lightchain further includes a Kappa variable region. In some embodiments, thefirst light chain further includes a Lambda variable region. In someembodiments, the second light chain includes at least a Lambda constantregion. In some embodiments, the second light chain further includes aLambda variable region. In some embodiments, the second light chainfurther includes a Kappa variable region. In some embodiments, the firstlight chain includes a Kappa constant region and a Kappa variableregion, and wherein the second light chain includes a Lambda constantregion and a Lambda variable region.

In some embodiments, the constant and variable framework regionsequences are human.

The invention also provides monovalent antibodies that bind CD47. Theseantibodies are collectively referred to herein as anti-CD47 monovalentantibodies or anti-CD47 monov mAbs. The monovalent antibodies of theinvention include one arm that specific recognizes CD47, and a secondarm referred to herein as a dummy arm. The dummy arm includes an aminoacid sequence that does not bind or otherwise cross-react with a humanprotein. In some embodiments, the dummy arm includes an amino acidsequence that does not bind or otherwise cross-react with a humanprotein that is found in whole blood. Those of ordinary skill in the artwill appreciate that human proteins found in the blood are a proxy thatrepresent all, or substantially all, antigens present in systemcirculation. In some embodiments, the dummy arm includes an amino acidsequence that does not bind or otherwise cross-react with a humanprotein that is found in solid tissue. Preferably, the monovalentantibodies are specific for at least human CD47. In some embodiments,the monovalent antibodies that recognize human CD47 are alsocross-reactive for at least one other non-human CD47 protein, such as,by way of non-limiting example, non-human primate CD47, e.g., cynomolgusmonkey CD47, and/or rodent CD47.

The anti-CD47 arms of the monovalent antibodies of the invention areuseful with any dummy arm. Exemplary anti-CD47 arms of the monovalentantibodies of the invention include the antibodies referred to herein asthe 5A3 antibody, the 5A3M4 antibody, the 5A3M3 antibody, the 5A3M5antibody, the KE8 antibody, the KE8-P6H5 antibody (also referred toherein as KE8H5), the KE8-P3B2 antibody (also referred to herein asKE8B2), the KE8-P2A2 antibody (also referred to herein as KE8A25), theKE8F2 antibody, the KE8G2 antibody, the KE84G9 antibody, the KE81G9antibody, the KE81A3 antibody, the KE8E8 antibody, the KE8G6 antibody,the KE8H3 antibody, the KE8C7 antibody, the KE8A4 antibody, the KE8A8antibody, the KE8G11 antibody, the KE8B7 antibody, the KE8F1 antibody,the KE8C4 antibody, the KE8A3 antibody, the KE86G9 antibody, the KE8H6antibody, the KA3 antibody, the KA3-P5G2 antibody (also referred toherein as KA3G2), the KA3-P1A3 antibody (also referred to herein asKA3A3), the KA3-P5C5 antibody (also referred to herein as KA3C5), theKA3H8 antibody, the KA3B2 antibody, the KA3A2 antibody, the KA3D3antibody, the KA3H3 antibody, the KC4 antibody, the KC4-P1G11KC4-P4C11antibody, the KC4-P6B1KC4-P4F4 antibody, and the KC4-P2E2 antibody (alsoreferred to herein as KC4E2), the KC4 antibody, the KC4F4 antibody, theKC4A1 antibody, the KC4C11 antibody, the KC4E10 antibody, the KC4B1antibody, the KC4C3 antibody, the KC4A4 antibody, the KC4G11 antibody,the KC4G9 antibody and fragments thereof. In some embodiments, theTAA-binding arm of the bispecific antibody includes an anti-CD19antibody sequence or antigen-binding fragment thereof.

In some embodiments, the monovalent antibody inhibits interactionbetween human CD47 and human SIRPα.

In some embodiments, the anti-CD47 arm of the monovalent antibodyincludes a first amino acid sequence that includes a variable heavychain complementarity determining region 1 (CDRH1) amino acid sequenceof SEQ ID NO: 225, a variable heavy chain complementarity determiningregion 2 (CDRH2) amino acid sequence of SEQ ID NO: 226, a variable heavychain complementarity determining region 3 (CDRH3) amino acid sequenceof SEQ ID NO: 227, a variable light chain complementarity determiningregion 1 (CDRL1) amino acid sequence selected from SEQ ID NO: 228-241and 262-272, a variable light chain complementarity determining region 2(CDRL2) amino acid sequence selected from 242-245 and 273-280, and avariable light chain complementarity determining region 3 (CDRH3) aminoacid sequence selected from 246-261 and 281.

In some embodiments, the anti-CD47 arm of the monovalent antibodyincludes a first amino acid sequence that includes a variable heavychain amino acid sequence of SEQ ID NO: 114 and a variable light chainamino acid sequence selected from SEQ ID NO: 116, 118, 120, 122, 124,126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152,154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180,182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204 and 206.

In some embodiments, the monovalent antibody includes two copies of asingle heavy chain polypeptide and a first light chain and a secondlight chain, wherein the first and second light chains are different.

In some embodiments, at least a portion of the first light chain is ofthe Kappa type and at least a portion of the second light chain is ofthe Lambda type. In some embodiments, the first light chain includes atleast a Kappa constant region. In some embodiments, the first lightchain further includes a Kappa variable region. In some embodiments, thefirst light chain further includes a Lambda variable region. In someembodiments, the second light chain includes at least a Lambda constantregion. In some embodiments, the second light chain further includes aLambda variable region. In some embodiments, the second light chainfurther includes a Kappa variable region. In some embodiments, the firstlight chain includes a Kappa constant region and a Kappa variableregion, and wherein the second light chain includes a Lambda constantregion and a Lambda variable region.

In some embodiments, the constant and variable framework regionsequences are human.

The bispecific antibodies of the invention are generated using anymethods known in the art such as, by way of non-limiting example, theuse of cross-linked fragments, quadromas, and/or any of a variety ofrecombinant formats such as, by way of non-limiting examples, linkedantibody fragments, forced heterodimers, and or recombinant formatsbased on single domains. Examples of Bispecific formats include but arenot limited to bispecific IgG based on Fab arm exchange (Gramer et al.,2013 MAbs. 5(6)); the CrossMab format (Klein C et al., 2012 MAbs 4(6));multiple formats based on forced heterodimerization approaches such asSEED technology (Davis J H et al., 2010 Protein Eng Des Sel.23(4):195-202), electrostatic steering (Gunasekaran K et al., J BiolChem. 2010 285(25):19637-46.) or knob-into-hole (Ridgway J B et al.,Protein Eng. 1996 9(7):617-21.) or other sets of mutations preventinghomodimer formation (Von Kreudenstein T S et al., 2013 MAbs.5(5):646-54.); fragment based bispecific formats such as tandem scFv(such asBiTEs) (Wolf E et al., 2005 Drug Discov. Today 10(18):1237-44.);bispecific tetravalent antibodies (Portner L M et al., 2012 CancerImmunol Immunother. 61(10):1869-75.); dual affinity retargetingmolecules (Moore P A et al., 2011 Blood. 117(17):4542-51), diabodies(Kontermann R E et al., Nat Biotechnol. 1997 15(7):629-31).

In some embodiments, the bispecific antibodies carry a differentspecificity in each combining site and including two copies of a singleheavy chain polypeptide and a first light chain and a second lightchain, wherein the first and second light chains are different.

In some antibodies, at least a first portion of the first light chain isof the Kappa type and at least a portion of the second light chain is ofthe Lambda type. In some antibodies, the first light chain includes atleast a Kappa constant region. In some antibodies, the first light chainfurther includes a Kappa variable region. In some antibodies, the firstlight chain further includes a Lambda variable region. In someantibodies, the second light chain includes at least a Lambda constantregion. In some antibodies, the second light chain further includes aLambda variable region. In some antibodies, the second light chainfurther includes a Kappa variable region. In some antibodies, the firstlight chain includes a Kappa constant region and a Kappa variableregion, and the second light chain includes a Lambda constant region anda Lambda variable region. In some embodiments, the constant and variableframework region sequences are human.

These anti-CD47 arms, monospecific anti-CD47 antibodies, monovalentanti-CD47 antibodies, and/or bispecific antibodies in which at least onebinding site is specific for CD47 contain a variable heavy chain aminoacid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or more identical to the amino acid sequence of SEQ ID NO: 114and a variable light chain amino acid sequence that is at least 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to anamino acid sequence selected from SEQ ID NO: 116, 118, 120, 122, 124,126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152,154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180,182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204 and 206.

The invention provides monoclonal antibodies that bind CD47. Forexample, the invention provides monoclonal antibodies that inhibit theinteraction between human CD47 at the surface of cells and soluble humanSIRPα with an IC50 greater than 0.3 nM in the assay described in Example4 and in which the antibody 5A3M3 has an IC50 of approximately 0.36 nM.

The invention also provides monoclonal antibodies that bind CD47 and arerecovered at more than 80% after incubation at 37° C. for 30 minutes inhuman whole blood at a concentration of 10 μg/ml as described in Example15. In some embodiments, the monoclonal antibody inhibits interactionbetween CD47 and signal-regulatory protein alpha (SIRPα). In someembodiments, the monoclonal antibody inhibits interaction between humanCD47 and human SIRPα.

The invention also provides anti-CD47 monoclonal antibodies that includea variable heavy chain complementarity determining region 1 (CDRH1)amino acid sequence of SEQ ID NO: 225, a variable heavy chaincomplementarity determining region 2 (CDRH2) amino acid sequence of SEQID NO: 226, a variable heavy chain complementarity determining region 3(CDRH3) amino acid sequence of SEQ ID NO: 227, a variable light chaincomplementarity determining region 1 (CDRL1) amino acid sequenceselected from SEQ ID NO: 228-241 and 262-272, a variable light chaincomplementarity determining region 2 (CDRL2) amino acid sequenceselected from 242-245 and 273-280, and a variable light chaincomplementarity determining region 3 (CDRH3) amino acid sequenceselected from 246-261 and 281.

In some embodiments, the anti-CD47 monoclonal antibody includes avariable heavy chain amino acid sequence that is at least 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acidsequence of SEQ ID NO: 114. In some embodiments, the anti-CD47monoclonal antibody includes a variable light chain amino acid sequencethat is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% ormore identical to an amino acid sequence selected from SEQ ID NO: 116,118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144,146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172,174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200,202, 204 and 206. In some embodiments, the anti-CD47 monoclonal antibodyincludes a variable heavy chain amino acid sequence that is at least90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical tothe amino acid sequence of SEQ ID NO: 114, and a variable light chainamino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or more identical to an amino acid sequence selected fromSEQ ID NO: 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138,140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166,168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194,196, 198, 200, 202, 204 and 206.

In some embodiments, the anti-CD47 monoclonal antibody includes avariable heavy chain amino acid sequence of SEQ ID NO: 114 and avariable light chain amino acid sequence selected from SEQ ID NO: 116,118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144,146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172,174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200,202, 204 and 206.

In some embodiments, the anti-CD47 antibody includes a combination of avariable heavy chain sequence and a variable light chain sequenceselected from the group consisting of the combinations shown in 5A3,5A3M4, 5A3M3, 5A3M5, KE8, KE8F2, KE8G2, KE84G9, KE81G9, KE81A3, KE8E8,KE8G6, KE8H5, KE8A2, KE8H3, KE8C7, KE8B2, KE8A4, KE8A8, KE8G11, KE8B7,KE8F1, KE8C4, KE8A3, KE86G9, KE8H6, KA3, KA3H8, KA3A3, KA3C5, KA3B2,KA3A2, KA3D3, KA3G2, KA3H3, KC4, KC4E2, KC4F4, KC4A1, KC4C11, KC4E10,KC4B1, KC4C3, KC4A4, KC4G11, and KC4G9.

The invention provides monoclonal antibodies that bind CD19. Theseantibodies are collectively referred to herein as anti-CD19 monoclonalantibodies or anti-CD19 mAbs. Preferably, the monoclonal antibodies arespecific for at least human CD19. In some embodiments, the monoclonalantibodies that recognize human CD19 are also cross-reactive for atleast one other non-human CD19 protein, such as, by way of non-limitingexample, non-human primate CD19, e.g., cynomolgus monkey CD19, and/orrodent CD19.

In some embodiments, the anti-CD19 monoclonal antibody includes avariable heavy chain amino acid sequence that is at least 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acidsequence of SEQ ID NO: 114. In some embodiments, the anti-CD19monoclonal antibody includes a variable light chain amino acid sequencethat is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% ormore identical to an amino acid sequence selected from SEQ ID NO: 208,210, 212, 214, 216, 218, and 220. In some embodiments, the anti-CD19monoclonal antibody includes a variable heavy chain amino acid sequencethat is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% ormore identical to the amino acid sequence of SEQ ID NO: 114, and avariable light chain amino acid sequence that is at least 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to an amino acidsequence selected from SEQ ID NO: 208, 210, 212, 214, 216, 218, and 220.

In some embodiments, the anti-CD19 monoclonal antibody includes avariable heavy chain amino acid sequence of SEQ ID NO: 114 and avariable light chain amino acid sequence selected from SEQ ID NO: 208,210, 212, 214, 216, 218, and 220.

The invention also provides monovalent antibodies that bind CD19. Theseantibodies are collectively referred to herein as anti-CD19 monovalentantibodies or anti-CD19 monov mAbs. The monovalent antibodies of theinvention include one arm that specific recognizes CD19, and a secondarm referred to herein as a dummy arm. The dummy arm includes an aminoacid sequence that does not bind or otherwise cross-react with a humanprotein. In some embodiments, the dummy arm includes an amino acidsequence that does not bind or otherwise cross-react with a humanprotein that is found in whole blood. In some embodiments, the dummy armincludes an amino acid sequence that does not bind or otherwisecross-react with a human protein that is found in solid tissue.Preferably, the monovalent antibodies are specific for at least humanCD19. In some embodiments, the monovalent antibodies that recognizehuman CD19 are also cross-reactive for at least one other non-human CD19protein, such as, by way of non-limiting example, non-human primateCD19, e.g., cynomolgus monkey CD19, and/or rodent CD19.

The invention also provides bispecific antibodies that recognize CD19and a second target. In some embodiments, the second target is anantigen known to be associated or otherwise implicated in autoimmunediseases and/or inflammatory diseases, such as, for example, B-cellmediated autoimmune diseases and/or inflammatory diseases, including byway of non-limiting example, systemic lupus erythematosus (SLE),rheumatoid arthritis (RA), idiopathic thrombocytopenic purpura (ITP),Waldenstrom's hypergammaglobulinaemia, Sjogren's syndrome, multiplesclerosis (MS), and/or lupus nephritis.

In some embodiments, suitable second targets include, by way ofnon-limiting example, CD20, CD22, CD40, BAFFR, CD5, CD32b, ICOSL, IL6R,and/or IL21R.

The bispecific antibodies of the invention that recognize CD19 and asecond target are generated using any methods known in the art such as,by way of non-limiting example, the use of cross-linked fragments,quadromas, and/or any of a variety of recombinant formats such as, byway of non-limiting examples, linked antibody fragments, forcedheterodimers, and or recombinant formats based on single domains. Theinvention allows for the identification, production and purification ofbispecific antibodies that are undistinguishable in sequence fromstandard antibodies and where one of the binding sites is specific forCD19 and the second binding site is specific for another target, forexample a tumor-associated antigen (TAA). The unmodified nature of theantibodies of the invention provides them with favorable manufacturingand biochemical characteristics similar to standard monoclonalantibodies.

In some embodiments, the bispecific antibodies carry a differentspecificity in each combining site and including two copies of a singleheavy chain polypeptide and a first light chain and a second lightchain, wherein the first and second light chains are different.

In some antibodies, at least a first portion of the first light chain isof the Kappa type and at least a portion of the second light chain is ofthe Lambda type. In some antibodies, the first light chain includes atleast a Kappa constant region. In some antibodies, the first light chainfurther includes a Kappa variable region. In some antibodies, the firstlight chain further includes a Lambda variable region. In someantibodies, the second light chain includes at least a Lambda constantregion. In some antibodies, the second light chain further includes aLambda variable region. In some antibodies, the second light chainfurther includes a Kappa variable region. In some antibodies, the firstlight chain includes a Kappa constant region and a Kappa variableregion, and the second light chain includes a Lambda constant region anda Lambda variable region. In some embodiments, the constant and variableframework region sequences are human.

The monoclonal, monovalent and/or bispecific antibodies of the inventioncan be used for therapeutic intervention or as a research or diagnosticreagent. For example, the monoclonal, monovalent and/or bispecificantibodies of the invention are useful in methods of treating,preventing and/or delaying the progression of pathologies associatedwith aberrant CD47 and/or aberrant CD47-SIRPα expression and/or activityor alleviating a symptom associated with such pathologies, byadministering an antibody of the invention to a subject in which suchtreatment or prevention is desired. The subject to be treated is, e.g.,human. The monoclonal, monovalent and/or bispecific antibody isadministered in an amount sufficient to treat, prevent, delay theprogression or alleviate a symptom associated with the pathology.

In some embodiments, the monoclonal, monovalent and/or bispecificantibodies described herein are used in conjunction with one or moreadditional agents or a combination of additional agents. Suitableadditional agents include current pharmaceutical and/or surgicaltherapies for an intended application, such as, for example, cancer,inflammation and/or autoimmune diseases. In some embodiments, themonoclonal, monovalent and/or bispecific antibodies can be used inconjunction with rituximab.

In some embodiments, the monoclonal, monovalent and/or bispecificantibodies and the additional agent are formulated into a singletherapeutic composition, and the monoclonal, monovalent and/orbispecific antibody and additional agent are administeredsimultaneously. Alternatively, the ac monoclonal, monovalent and/orbispecific antibodies and additional agent are separate from each other,e.g., each is formulated into a separate therapeutic composition, andthe monoclonal, monovalent and/or bispecific antibody and the additionalagent are administered simultaneously, or the monoclonal, monovalentand/or bispecific antibodies and the additional agent are administeredat different times during a treatment regimen. For example, themonoclonal, monovalent and/or bispecific antibody is administered priorto the administration of the additional agent, the monoclonal,monovalent and/or bispecific antibody is administered subsequent to theadministration of the additional agent, or the monoclonal, monovalentand/or bispecific antibody and the additional agent are administered inan alternating fashion. As described herein, the monoclonal, monovalentand/or bispecific antibody and additional agent are administered insingle doses or in multiple doses.

Pathologies treated and/or prevented using the antibodies of theinvention include, for example, cancer or any other disease or disorderassociated with aberrant CD47 expression and/or activity.

The invention also provides methods of producing bispecific antibodiesthat exhibit an “unbalanced” affinity for each of the two targets. Forexample, in some embodiments of an anti-CD47/CD19 bispecific antibody,the affinity of the anti-CD19 arm is increased using affinitymaturation. For example, in some embodiments of an anti-CD47/CD19bispecific antibody, the affinity of the anti-CD47 arm is decreasedusing affinity dematuration. For example, in some embodiments ananti-CD47/CD19 bispecific antibody, the affinity of the anti-CD19 arm isincreased using affinity maturation, and the affinity of the anti-CD47arm is decreased using affinity de-maturation. These unbalanced affinitybispecific antibodies are useful, for example, to improve selectivityfor a target cell or group of target cells.

FIG. 1 is an illustration of the sequence alignment between the variablelight chain (VL) sequence of the anti-CD47 antibody 5A3 (SEQ ID NO: 285)to its closest germline sequence (SEQ ID NO: 282), the human IGKV1-33according to the IMGT nomenclature, and to the variable light chain (VL)sequence of the anti-CD47 antibody 5A3-M3-VL (SEQ ID NO: 286) and to thevariable light chain (VL) sequence of the anti-CD47 antibody 5A3-M5-VL(SEQ ID NO: 287)).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an illustration of the sequence alignment between the variablelight chain (VL) sequence of the anti-CD47 antibody 5A3 (SEQ ID NO: 116)to its closest germline sequence (SEQ ID NO: 282), the human IGKV1-33according to the IMGT nomenclature.

FIG. 2 is a graph depicting the blocking potency of the 5A3-M3 and5A3-M5 antibody variants as compared to the parental antibody 5A3 in aCD47/SIRPα binding assay.

FIG. 3 is a graph depicting the specificity of various CD47 monoclonalantibodies (Mabs) as indicated by the binding of purified CD47 Mabs toCHO cells transfected with human CD47, assessed by flow cytometry (greybars). CD47 MAbs did not bind to non-transfected CHO cells (black bars).

FIG. 4 is a graph depicting binding to native CD47 and specificity ofCD47 MAbs as shown by binding of purified CD47 Mabs to HEK293-P cells asassessed by flow cytometry (grey bars). Binding to HEK293-P cells stablytransfected with hCD47-specific siRNA is significantly decreased (blackbars).

FIG. 5 is a graph depicting binding to native human CD47 andcross-reactivity with cynomolgus CD47. The binding of purified CD47 Mabsto human (light grey bars) and cynomolgus (dark grey bars) PBMC CD4+ Tcells was evaluated.

FIG. 6 is a graph depicting the potency of CD47 Mabs to block theCD47-SIRPα interaction, as tested in the CD47-SIRPα inhibition assay(competitive inhibition of the binding of recombinant soluble humanSIRPα to hCD47-expressing CHO cells, as described in Example 4). IC50values obtained in dose-response experiments are shown. CD47 Mabs aregrouped by family and ranked from higher to lower potency. Theneutralizing activity of the antibodies of the present invention iscompared to the commercially available CD47 antibody B6H12.

FIG. 7 is an illustration depicting the hemagglutination activity ofCD47 antibodies. Hemagglutination is evidenced as a clumped deposit ofRBC, in the form of a crescent at the bottom around the inferior borderof the well, whereas non-agglutinated are do not form aggregates and aredistributed evenly over the well surface area. FIG. 7 demonstrates thathigh-affinity CD47 Mabs of the 5A3, Ke8, and Ka3 families inducehemagglutination; in contrast to the other three families, Kc4 familyantibodies tested in this experiment do not induce hemagglutination.

FIG. 8 is a graph depicting dose-response curve for a FACS-binding assaydone with Raji cells and the original anti-CD19 clone 1B7, clone D11identified following the first affinity maturation round, and the finalclone L7B7_c2, issued form the second affinity maturation round.

FIGS. 9A-9C are a series of graphs the ability of a CD47xCD19 BsAb toco-engage the two targets at the surface of cells. The graphs in FIGS.9A-9C present FACS profiles generated with monovalent and bispecificantibodies binding to CD19-negative B-NHL cells (DS-1) and CD19-positiveBurkitt lymphoma cells (Raji). All antibodies were human IgG1 format andwere tested at four concentrations as indicated.

FIG. 10 is a series of graphs depicting the SIRPα Blocking Activity ofMonovalent and Bispecific Antibodies. FIG. 10 demonstrates theco-engagement of CD19 and CD47 on the surface of the target cell, byshowing that the neutralization of CD47-SIRPα interaction by CD47xCD19BsAbs is CD19-dependent. The experiments were done in quadruplicatesMean and SEM are shown. Dose-response inhibition curves were fitted withGraphPad software.

FIGS. 11A-11C show ADCC dose-response curves generated with CD47xCD19 κλbodies (black) or the corresponding CD47 monovalent antibodies (grey).ADCC with the CD19 Mab C2 is shown for comparison (dashed grey). TheADCC assay was performed with whole human PBMCs as effector cells andCalcein AM-stained Raji (FIG. 11A, 11C) or Ramos (FIG. 11B) as targetcells (effector to target ratio: 50). Cytotoxicity was calculated fromthe degree of calcein release from target cells. The percentage ofspecific cell killing+/−SD is shown. The experiments were done induplicates. FIGS. 11A-11B demonstrate the ability of CD47xCD19 BsAbs tokill CD19-positive cells in a CD19-dependent manner, as thecorresponding CD47 monovalent antibodies were much less efficient or notefficient at all. FIG. 11C demonstrates that the efficacy of killing ofRaji cells with CD47xCD19 antibodies was comparable to rituximab andmuch higher than with the CD19 Mab C2.

FIG. 12 is a graph depicting the phagocytic activity of three of theCD47xCD19 κλ bodies of the present invention (black lines) compared tothe corresponding CD47 monovalent antibodies (grey lines) indose-response ADCP experiment. Phagocytosis with the CD47 Mab B6H12 (onhuman IgG1 background, dotted black line) and with the CD19 Mab C2(dotted grey line) is shown for comparison. The ADCP experiment wasperformed with human macrophages differentiated from peripheral bloodmonocytes and Raji as target cells (effector: target ratio 1:5)Phagocytosis was assessed by FACS. The percentage of macrophages havingphagocytosed at least one target cells is shown. CD47xCD19 κλ bodiesphagocytose CD19-positive cells in a CD19-dependent manner analysis, asthe corresponding CD47 monovalent antibodies were much less efficient ornot efficient at all.

FIG. 13 is a graph depicting the activity of various antibodies in aRaji B cell lymphoma xenograft in NOD/SCID mice. Antibody treatmentstarted after the tumor graft has reached about 0.1 cm³ and ended onD25. Treatment groups (n=5) were as indicated in the inset. Shown is theevolution of average tumor volume per treatment group+/−SD. FIG. 13shows that the efficacy of BsAB is similar to B6H12 or rituximab andthat tumor eradication was CD19-dependent, as the correspondingmonovalent was less efficacious.

FIG. 14 is a graph depicting that high and moderate affinity CD47antibodies are efficiently adsorbed on erythrocytes. In the case ofBsAbs, this phenomenon is limited to molecules having a high affinityCD47 arms, such as 5A3.

DETAILED DESCRIPTION

The invention provides monoclonal antibodies that bind CD47. Theseantibodies are collectively referred to herein as anti-CD47 monoclonalantibodies or anti-CD47 mAbs. Preferably, the monoclonal antibodies arespecific for at least human CD47. In some embodiments, the monoclonalantibodies that recognize human CD47 are also cross-reactive for atleast one other non-human CD47 protein, such as, by way of non-limitingexample, non-human primate CD47, e.g., cynomolgus monkey CD47, and/orrodent CD47. In some embodiments, these anti-CD47 monoclonal antibodiesinhibit the interaction between CD47 and signal-regulatory protein alpha(SIRPα). In some embodiments, these anti-CD47 monoclonal antibodiesinhibit the interaction between human CD47 and human SIRPα. Theinvention also include antibodies that bind to the same epitope as ananti-CD47 monoclonal antibody disclosed herein and inhibits theinteraction between CD47 and SIRPα, e.g., between human CD47 and humanSIRPα.

The invention also provides monovalent antibodies and/or bispecificantibodies that include at least a first arm that is specific for CD47.Preferably, the monovalent antibodies and/or bispecific antibodies arespecific for at least human CD47. In some embodiments, the monovalentantibodies and/or bispecific antibodies that recognize human CD47 arealso cross-reactive for at least one other non-human CD47 protein, suchas, by way of non-limiting example, non-human primate CD47, e.g.,cynomolgus monkey CD47, and/or rodent CD47. In some embodiments, theseanti-CD47 monovalent antibodies and/or anti-CD47 bispecific antibodiesinhibit the interaction between CD47 and signal-regulatory protein alpha(SIRPα). In some embodiments, these anti-CD47 monovalent antibodiesand/or anti-CD47 bispecific antibodies inhibit the interaction betweenhuman CD47 and human SIRPα. The invention also include antibodies thatbind to the same epitope as an anti-CD47 monovalent and/or an anti-CD47bispecific antibody disclosed herein and inhibits the interactionbetween CD47 and SIRPα, e.g., between human CD47 and human SIRPα.

The bispecific antibodies of the invention allow for simultaneousbinding of the two antibody arms to two antigens on the surface of thecell (termed co-engagement), which results in additive or synergisticincrease of affinity due to avidity mechanism. As a consequence,co-engagement confers high selectivity towards cells expressing bothantigens as compared to cells that express just one single antigen. Inaddition, the affinities of the two arms of a bispecific antibody totheir respective targets can be set up in a way that binding to targetcells is principally driven by one of the antibody arms. In someembodiments, the bispecific antibody includes a first arm that bindsCD47 and a second arm that binds a tumor associated antigen (TAA), wherethe second arm binds to the TAA with high affinity, and the first armbinds to CD47 with low affinity, i.e., an affinity that is sufficient toinhibit CD47/SIRPα upon TAA co-engagement. This design allows thebispecific antibodies of the invention to preferentially inhibit CD47 incancer versus normal cells. In the examples provided herein, abispecific antibody with a first arm that binds CD47 with low affinityand a second arm that binds CD19 with high affinity (termed a CD47xCD19bispecific) allow preferential inhibition of CD47 in cancer versusnormal cells. Besides the two antigen-binding arms, the CD47 x TAAbispecific antibody requires a functional Fc portion to recruitmacrophages and/or other immune effector cells. A fully human bispecificIgG format (such as the K-body format described herein) is well suitedfor the generation of dual targeting CD47 x TAA bispecific antibodies.As shown in the examples provided herein, the ability of dual targetingbispecific antibodies to co-engage CD47 and CD19 results in asignificant increase in the affinity of binding to CD19-positive cellsand in CD19-dependent neutralization of the CD47-SIRPα interaction.This, in turn, translates into efficient and selective cancer cellkilling mediated by the CD47xCD19 bispecific antibody, as demonstratedin the ADCC and ADCP experiments provided herein.

Exemplary anti-CD47 monoclonal, monospecific anti-CD47 antibodies,anti-CD47 monovalent antibodies, and/or bispecific antibodies of theinvention in which at least one binding site is specific for CD47include, for example, the 5A3 antibody, the 5A3M4 antibody, the 5A3M3antibody, the 5A3M5 antibody, the KE8 antibody, the KE8-P6H5 antibody(also referred to herein as KE8H5), the KE8-P3B2 antibody (also referredto herein as KE8B2), the KE8-P2A2 antibody (also referred to herein asKE8A25), the KE8F2 antibody, the KE8G2 antibody, the KE84G9 antibody,the KE81G9 antibody, the KE81A3 antibody, the KE8E8 antibody, the KE8G6antibody, the KE8H3 antibody, the KE8C7 antibody, the KE8A4 antibody,the KE8A8 antibody, the KE8G11 antibody, the KE8B7 antibody, the KE8F1antibody, the KE8C4 antibody, the KE8A3 antibody, the KE86G9 antibody,the KE8H6 antibody, the KA3 antibody, the KA3-P5G2 antibody (alsoreferred to herein as KA3G2), the KA3-P1A3 antibody (also referred toherein as KA3A3), the KA3-P5C5 antibody (also referred to herein asKA3C5), the KA3H8 antibody, the KA3B2 antibody, the KA3A2 antibody, theKA3D3 antibody, the KA3H3 antibody, the KC4 antibody, theKC4-P1G11KC4-P4C11 antibody, the KC4-P6B1KC4-P4F4 antibody, and theKC4-P2E2 antibody (also referred to herein as KC4E2), the KC4 antibody,the KC4F4 antibody, the KC4A1 antibody, the KC4C11 antibody, the KC4E10antibody, the KC4B1 antibody, the KC4C3 antibody, the KC4A4 antibody,the KC4G11 antibody, and the KC4G9 antibody, as well as immunologicallyactive and/or antigen-binding fragments thereof.

In some embodiments, exemplary anti-CD47 monoclonal, monospecificanti-CD47 antibodies, anti-CD47 monovalent antibodies, and/or bispecificantibodies of the invention include a combination of heavy chain andlight chain complementarity determining regions (CDRs) selected from theCDR sequences shown in Tables 1, 2 and 3, where the CDRs shown in Tables1, 2 and 3 are defined according to the IMGT nomenclature.

In some embodiments, exemplary anti-CD47 monoclonal, monospecificanti-CD47 antibodies, anti-CD47 monovalent antibodies, and/or bispecificantibodies of the invention include the combination of heavy chain CDRsequences from Table 1 and two sets of light chain CDRs selected fromthe CDRL1, CDRL2 and CDRL3 sequences shown in Tables 2 and 3.

In some embodiments, exemplary anti-CD47 monoclonal, monospecificanti-CD47 antibodies, anti-CD47 monovalent antibodies, and/or bispecificantibodies of the invention include the combination of heavy chain CDRsequences from Table 1 and a first set of light chain CDRs selected fromthe CDRL1, CDRL2 and CDRL3 sequences shown in Table 2 and a second setof light chain CDRs selected from the CDRL1, CDRL2 and CDRL3 sequencesshown in Table 3.

TABLE 1 Anti-CD47 Heavy Chain CDRs CDRH1 CDRH2 CDRH3 GFTF----SSYAISGS--GGST AKSYGAF----DY (SEQ ID NO: 225) (SEQ ID NO: 226) (SEQ ID NO:227)

TABLE 2 Anti-CD47 Kappa Light Chain CDRs CDRL1 CDRL2 CDRL3 QDI------NKYAA-------S QQKHPRGP---RT (SEQ ID NO: 228) (SEQ ID NO: 242) (SEQ ID NO:246) QDI------NRY GA-------S QQFHKRAP---QT (SEQ ID NO: 229) (SEQ ID NO:243) (SEQ ID NO: 247) QNI------GKY NA-------S QQFHKRRP---QT (SEQ ID NO:230) (SEQ ID NO: 244) (SEQ ID NO: 248) QSI------ARY SA-------SQQFHKRSP---QT (SEQ ID NO: 231) (SEQ ID NO: 245) (SEQ ID NO: 249)QSI------ASY QQKHPRAP---RT (SEQ ID NO: 232) (SEQ ID NO: 250)QSI------DKY QQKHPRSP---RT (SEQ ID NO: 233) (SEQ ID NO: 251)QSI------DRY QQKHPRYP---RT (SEQ ID NO: 234) (SEQ ID NO: 252)QSI------GKY QQKHPRNP---RT (SEQ ID NO: 235) (SEQ ID NO: 253)QSI------GRY QQMHPRAP---KT (SEQ ID NO: 236) (SEQ ID NO: 254)QSI------NRY QQMHPRGP---KT (SEQ ID NO: 237) (SEQ ID NO: 255)QSI------SKY QQMHPRSP---KT (SEQ ID NO: 238) (SEQ ID NO: 256)QSI------SRY QQRHPRAP---RT (SEQ ID NO: 239) (SEQ ID NO: 257)QSI------SSY QQRHKRSP---QT (SEQ ID NO: 240) (SEQ ID NO: 258)QSI------AKY QQRHPRGP---RT (SEQ ID NO: 241) (SEQ ID NO: 259)QQRHPRGP---ST (SEQ ID NO: 260) QQRHPRGP---TT (SEQ ID NO: 261)

TABLE 3 Anti-CD47 Lambda Light Chain CDRs CDRL1 CDRL2 CDRL3 SSDVG---GYNYEN-------S SSYDWWFRP--KV (SEQ ID NO: 262) (SEQ ID NO: 273) (SEQ ID NO:281) SSDVE---RKNY ES-------S (SEQ ID NO: 263) (SEQ ID NO: 274)SSDVR---ANNY EV-------S (SEQ ID NO: 264) (SEQ ID NO: 275) SSDVY---YNKYKD-------S (SEQ ID NO: 265) (SEQ ID NO: 276) SSDVG---KANY KN-------S(SEQ ID NO: 266) (SEQ ID NO: 277) SSDVR---GNNY KS-------S (SEQ ID NO:267) (SEQ ID NO: 278) SSDVS---ARNY KT-------S (SEQ ID NO: 268) (SEQ IDNO: 279) SSDVN---SANY QD-------S (SEQ ID NO: 269) (SEQ ID NO: 280)SSDVR---AANY (SEQ ID NO: 270) SSDVR---RANY (SEQ ID NO: 271) SSDVN---NTNY(SEQ ID NO: 272)

Each of the exemplary anti-CD47, anti-CD19, monovalent and bispecificantibodies described herein include a common heavy chain (HC), one kappachain or one lambda chain for anti-CD47 and anti-CD19 antibodies, onekappa and one lambda light chains (LC) for monovalent and bispecificantibodies, as shown in the amino acid and corresponding nucleic acidsequences listed below. Each of the exemplary anti-CD47, anti-CD19,monovalent and bispecific antibodies described below includes a commonvariable heavy domain (VH), one kappa variable light domain or onelambda variable light domain for anti-CD47 and anti-CD19 antibodies, onekappa and one lambda variable light domains (VL) for monovalent andbispecific antibodies, as shown in the amino acid and correspondingnucleic acid sequences listed below.

While antibody sequences below are provided herein as examples, it is tobe understood that these sequences can be used to generate bispecificantibodies using any of a variety of art-recognized techniques. Examplesof bispecific formats include but are not limited to bispecific IgGbased on Fab arm exchange (Gramer et al., 2013 MAbs. 5(6)); the CrossMabformat (Klein C et al., 2012 MAbs 4(6)); multiple formats based onforced heterodimerization approaches such as SEED technology (Davis J Het al., 2010 Protein Eng Des Sel. 23(4): 195-202), electrostaticsteering (Gunasekaran K et al., J Biol Chem. 2010 285(25):19637-46.) orknob-into-hole (Ridgway J B et al., Protein Eng. 1996 9(7):617-21.) orother sets of mutations preventing homodimer formation (Von KreudensteinT S et al., 2013 MAbs. 5(5):646-54.); fragment based bispecific formatssuch as tandem scFv (such asBiTEs) (Wolf E et al., 2005 Drug Discov.Today 10(18):1237-44.); bispecific tetravalent antibodies (Portner L Met al., 2012 Cancer Immunol Immunother. 61(10):1869-75.); dual affinityretargeting molecules (Moore P A et al., 2011 Blood. 117(17):4542-51),diabodies (Kontermann R E et al., Nat Biotechnol. 1997 15(7):629-31).

The exemplary anti-CD47, anti-CD19, monovalent and bispecific antibodiesinclude a common heavy chain (SEQ ID NO: 2) encoded by the nucleic acidsequence shown in SEQ ID NO: 1.

>COMMON-HC-NT (SEQ ID NO: 1)GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAAGTTATGGTGCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACAGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACAGTCTCGTGGAACTCAGGAGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACTGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCTCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACTTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGTCCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTTAA >COMMON-HC-AA (SEQ ID NO: 2)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSYGAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

The anti-CD47, anti-CD19, monovalent and bispecific antibodies include acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113.

>COMMON-VH-NT (SEQ ID NO: 113)GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAAGTTATGGTGCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACAGTCTCGAGC >COMMON-VH-AA (SEQ IDNO: 114) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSY GAFDYWGQGTLVTVSS

Anti-CD47 Antibodies

The 5A3 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 4) encoded by the nucleic acid sequence shown inSEQ ID NO: 3.

>5A3-LC-NT (SEQ ID NO: 3)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >5A3-LC-AA (SEQ ID NO:4) DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The 5A3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 116) encoded by thenucleic acid sequence shown in SEQ ID NO: 115.

>5A3-VL-NT (SEQ ID NO: 115)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >5A3-VL-AA (SEQ ID NO: 116)DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRGPRTFG QGTKVEIK

The 5A3-M4 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 6) encoded by the nucleic acid sequence shown inSEQ ID NO: 5.

>5A3-M4-LC-NT (SEQ ID NO: 5)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGAACCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >5A3-M4-LC-AA (SEQ IDNO: 6) DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRNPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The 5A3-M4 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 118) encoded by thenucleic acid sequence shown in SEQ ID NO: 117.

>5A3-M4-VL-NT (SEQ ID NO: 117)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGAACCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >5A3-M4-VL-AA (SEQ ID NO: 118)DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRNPRTFG QGTKVEIK

The 5A3-M3 antibody includes a common heavy chain (SEQ TD NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 8) encoded by the nucleic acid sequence shown inSEQ ID NO: 7.

>5A3-M3-LC-NT (SEQ ID NO: 7)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGTCCATTAGTAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGCTGCATCCTCGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >5A3-M3-LC-AA (SEQ IDNO: 8) DIQMTQSPSSLSASVGDRVTITCQASQSISSYLNWYQQKPGKAPKLLIYAASSLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The 5A3-M3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ TD NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 120) encoded by thenucleic acid sequence shown in SEQ ID NO: 119.

>5A3-M3-VL-NT (SEQ ID NO: 119)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGTCCATTAGTAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGCTGCATCCTCGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >5A3-M3-VL-AA (SEQ ID NO: 120)DIQMTQSPSSLSASVGDRVTITCQASQSISSYLNWYQQKPGKAPKLLIYAASSLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRGPRTFG QGTKVEIK

The 5A3-M5 antibody includes a common heavy chain (SEQ TD NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 10) encoded by the nucleic acid sequence shownin SEQ TD NO: 9.

>5A3-M5-LC-NT (SEQ ID NO: 9)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGTACCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >5A3-M5-LC-AA (SEQ IDNO: 10) DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRYPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The 5A3-M5 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 122) encoded by thenucleic acid sequence shown in SEQ ID NO: 121.

>5A3-M5-VL-NT (SEQ ID NO: 121)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGGTGCATCCAGGTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGCAGAAGCACCCCCGGTACCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >5A3-M5-VL-A (SEQ ID NO: 122)DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYGASRLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQKHPRYPRTFG QGTKVEIK

The Ke8 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 12) encoded by the nucleic acid sequence shownin SEQ ID NO: 11.

>Ke8-LC-NT (SEQ ID NO: 11)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCACAAGCGGCGGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8-LC-AA (SEQ ID NO:12) DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRRPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 124) encoded by thenucleic acid sequence shown in SEQ ID NO: 123.

>Ke8-VL-NT (SEQ ID NO: 123)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCACAAGCGGCGGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8-VL-AA (SEQ ID NO: 124)DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRRPQTFG QGTKVEIK

The Ke8H5 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 14) encoded by the nucleic acid sequence shownin SEQ ID NO: 13.

>KE8H5-LC-NT (SEQ ID NO: 13)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCGAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTGCGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8H5-LC-AA (SEQ IDNO: 14) DIQMTQSPSSLSASVGDRVTITCRASQSIARYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRAPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8H5 antibody includes a common variable heavy domain (SEQ TD NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 126) encoded by thenucleic acid sequence shown in SEQ ID NO: 125.

>KE8H5-VL-NT (SEQ ID NO: 125)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCGAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTGCGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8H5-VL-AA (SEQ ID NO: 126)DIQMTQSPSSLSASVGDRVTITCRASQSIARYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRAPQTFG QGTKVEIK

The Ke8B2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 16) encoded by the nucleic acid sequence shownin SEQ ID NO: 15.

>KE8B2-LC-NT (SEQ ID NO: 15)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCACCCGCGTGCCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8B2-LC-AA (SEQ IDNO: 16) DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRAPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8B2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 128) encoded by thenucleic acid sequence shown in SEQ ID NO: 127.

>KE8B2-VL-NT (SEQ ID NO: 127)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCACCCGCGTGCCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8B2-VL-AA (SEQ ID NO: 128)DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRAPRTFG QGTKVEIK

The Ke8A2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 18) encoded by the nucleic acid sequence shownin SEQ ID NO: 17.

>KE8A2-LC-NT (SEQ ID NO: 17)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8A2-LC-AA (SEQ IDNO: 18) DIQMTQSPSSLSASVGDRVTITCRASQSIDRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8A2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 130) encoded by thenucleic acid sequence shown in SEQ ID NO: 129.

>KE8A2-VL-NT (SEQ ID NO: 129)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8A2-VL-AA (SEQ ID NO: 130)DIQMTQSPSSLSASVGDRVTITCRASQSIDRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFG QGTKVEIK

The Ke8E8 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 20) encoded by the nucleic acid sequence shownin SEQ ID NO: 19.

>KE8E8-LC-NT (SEQ ID NO: 19)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGGCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8E8-LC-AA (SEQ IDNO: 20) DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8E8 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 132) encoded by thenucleic acid sequence shown in SEQ ID NO: 131.

>KE8E8-VL-NT (SEQ ID NO: 131)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGGCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8E8-VL-AA (SEQ ID NO: 132)DIQMTQSPSSLSASVGDRVTITCQASQDINKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFG QGTKVEIK

The Ke8H3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 22) encoded by the nucleic acid sequence shownin SEQ TD NO: 21.

>KE8H3-LC-NT (SEQ ID NO: 21)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8H3-LC-AA (SEQ IDNO: 22) DIQMTQSPSSLSASVGDRVTITCRASQSINRYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8H3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 134) encoded by thenucleic acid sequence shown in SEQ ID NO: 133.

>KE8H3-VL-NT (SEQ ID NO: 133)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTGGGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8H3-VL-AA (SEQ ID NO: 134)DIQMTQSPSSLSASVGDRVTITCRASQSINRYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRGPRTFG QGTKVEIK

The Ke8G6 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 24) encoded by the nucleic acid sequence shownin SEQ ID NO: 23.

>KE8G6-LC-NT (SEQ ID NO: 23)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGCGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8G6-LC-AA (SEQ IDNO: 24) DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8G6 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 136) encoded by thenucleic acid sequence shown in SEQ ID NO: 135.

>KE8G6-VL-NT (SEQ ID NO: 135)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGCGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8G6-VL-AA (SEQ ID NO: 136)DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ke8A3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 26) encoded by the nucleic acid sequence shownin SEQ ID NO: 25.

>KE8A3-LC-NT (SEQ ID NO: 25)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGTAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCCCGTGGGCCGAGCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8A3-LC-AA (SEQ IDNO: 26) DIQMTQSPSSLSASVGDRVTITCRVSQSISKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPSTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8A3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 138) encoded by thenucleic acid sequence shown in SEQ ID NO: 137.

>KE8A3-VL-NT (SEQ ID NO: 137)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGTAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCCCGTGGGCCGAGCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8A3-VL-AA (SEQ ID NO: 138)DIQMTQSPSSLSASVGDRVTITCRVSQSISKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPSTFG QGTKVEIK

The Ke81A3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 28) encoded by the nucleic acid sequence shownin SEQ ID NO: 27.

>KE81A3-LC-NT (SEQ ID NO: 27)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGCCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE81A3-LC-AA (SEQ IDNO: 28) DIQMTQSPSSLSASVGDRVTITCQASQDINRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRAPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke81A3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ TD NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 140) encoded by thenucleic acid sequence shown in SEQ ID NO: 139.

>KE81A3-VL-NT (SEQ ID NO: 139)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGCCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE81A3-VL-AA (SEQ ID NO: 140)DIQMTQSPSSLSASVGDRVTITCQASQDINRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRAPRTFG QGTKVEIK

The Ke8A8 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 30) encoded by the nucleic acid sequence shownin SEQ ID NO: 29.

>KE8A8-LC-NT (SEQ ID NO: 29)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGCGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8A8-LC-AA (SEQ IDNO: 30) DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8A8 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 142) encoded by thenucleic acid sequence shown in SEQ ID NO: 141.

>KE8A8-VL-NT (SEQ ID NO: 141)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGCGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8A8-VL-AA (SEQ ID NO: 142)DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ke8C7 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 32) encoded by the nucleic acid sequence shownin SEQ ID NO: 31.

>KE8C7-LC-NT (SEQ ID NO: 31)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGCGCCATCCGCGTGGCCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8C7-LC-AA (SEQ IDNO: 32) DIQMTQSPSSLSASVGDRVTITCQASQDINRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8C7 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 144) encoded by thenucleic acid sequence shown in SEQ ID NO: 143.

>KE8C7-VL-NT (SEQ ID NO: 143)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTAATAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGCGCCATCCGCGTGGCCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8C7-VL-AA (SEQ ID NO: 144)DIQMTQSPSSLSASVGDRVTITCQASQDINRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPRTFG QGTKVEIK

The Ke8G2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 34) encoded by the nucleic acid sequence shownin SEQ TD NO: 33.

>KE8G2-LC-NT (SEQ ID NO: 33)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGCGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8G2-LC-AA (SEQ IDNO: 34) DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTINSLQPEDFATYYCQQKHPRAPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8G2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 146) encoded by thenucleic acid sequence shown in SEQ ID NO: 145.

>KE8G2-VL-NT (SEQ ID NO: 145)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCCCGTGCGCCGAGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8G2-VL-AA (SEQ ID NO: 146)DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTINSLQPEDFATYYCQQKHPRAPRTFG QGTKVEIK

The Ke81G9 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 36) encoded by the nucleic acid sequence shownin SEQ ID NO: 35.

>KE81G9-LC-NT (SEQ ID NO: 35)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGCGGCATAAGCGTTCCCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE81G9-LC-AA (SEQ IDNO: 36) DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHKRSPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke81G9 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 148) encoded by thenucleic acid sequence shown in SEQ ID NO: 147.

>KE81G9-VL-NT (SEQ ID NO: 147)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGCGGCATAAGCGTTCCCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE81G9-VL-AA (SEQ ID NO: 148)DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHKRSPQTFG QGTKVEIK

The Ke8F2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 38) encoded by the nucleic acid sequence shownin SEQ ID NO: 37.

>KE8F2-LC-NT (SEQ ID NO: 37)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTGCGCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8F2-LC-AA (SEQ IDNO: 38) DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRAPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8F2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 150) encoded by thenucleic acid sequence shown in SEQ ID NO: 149.

>KE8F2-VL-NT (SEQ ID NO: 149)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTGCGCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8F2-VL-AA (SEQ ID NO: 150)DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRAPRTFG QGTKVEIK

The Ke8B7 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 40) encoded by the nucleic acid sequence shownin SEQ ID NO: 39.

>KE8B7-LC-NT (SEQ ID NO: 39)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGGAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTAGCCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8B7-LC-AA (SEQ IDNO: 40) DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8B7 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 152) encoded by thenucleic acid sequence shown in SEQ ID NO: 151.

>KE8B7-VL-NT (SEQ ID NO: 151)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGGAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTAGCCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8B7-VL-AA (SEQ ID NO: 152)DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFG QGTKVEIK

The Ke8C4 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 42) encoded by the nucleic acid sequence shownin SEQ ID NO: 41.

>KE8C4-LC-NT (SEQ ID NO: 41)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGGGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8C4-LC-AA (SEQ IDNO: 42) DIQMTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8C4 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 154) encoded by thenucleic acid sequence shown in SEQ ID NO: 153.

>KE8C4-VL-NT (SEQ ID NO: 153)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGGGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8C4-VL-AA (SEQ ID NO: 154)DIQMTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYAASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFG QGTKVEIK

The Ke8F1 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 44) encoded by the nucleic acid sequence shownin SEQ ID NO: 43.

>KE8F1-LC-NT (SEQ ID NO: 43)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTTCTTATGTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTCGGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8F1-LC-AA (SEQ IDNO: 44) DIQMTQSPSSLSASVGDRVTITCRASQSIASYVNWYQQKPGKAPKLLIYAASGLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRRPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8F1 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 156) encoded by thenucleic acid sequence shown in SEQ ID NO: 155.

>KE8F1-VL-NT (SEQ ID NO: 155)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTTCTTATGTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTCGGCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8F1-VL-AA (SEQ ID NO: 156)DIQMTQSPSSLSASVGDRVTITCRASQSIASYVNWYQQKPGKAPKLLIYAASGLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRRPQTFG QGTKVEIK

The Ke8G11 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 46) encoded by the nucleic acid sequence shownin SEQ ID NO: 45.

>KE8G11-LC-NT (SEQ ID NO: 45)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGGAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGGGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8G11-LC-AA (SEQ IDNO: 46) DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8G11 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 158) encoded by thenucleic acid sequence shown in SEQ ID NO: 157.

>KE8G11-VL-NT (SEQ ID NO: 157)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGGAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCGCGTGGGCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8G11-VL-AA (SEQ ID NO: 158)DIQMTQSPSSLSASVGDRVTITCRASQSIGRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFG QGTKVEIK

The Ke8H6 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 48) encoded by the nucleic acid sequence shownin SEQ TD NO: 47.

>KE8H6-LC-NT (SEQ ID NO: 47)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAATGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGGGCCGCGCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8H6-LC-AA (SEQ IDNO: 48) DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYNASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8H6 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 160) encoded by thenucleic acid sequence shown in SEQ ID NO: 159.

>KE8H6-VL-NT (SEQ ID NO: 159)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAATGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGGGCCGCGCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8H6-VL-AA (SEQ ID NO: 160)DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYNASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPRTFG QGTKVEIK

The Ke84G9 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 50) encoded by the nucleic acid sequence shownin SEQ ID NO: 49.

>KE84G9-LC-NT (SEQ ID NO: 49)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTAGCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE84G9-LC-AA (SEQ IDNO: 50) DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRSPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke84G9 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 162) encoded by thenucleic acid sequence shown in SEQ ID NO: 161.

>KE84G9-VL-NT (SEQ ID NO: 161)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAAGCATCCGCGTAGCCCGCGGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE84G9-VL-AA (SEQ ID NO: 162)DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQKHPRSPRTFG QGTKVEIK

The Ke8A4 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 52) encoded by the nucleic acid sequence shownin SEQ ID NO: 51.

>KE8A4-LC-NT (SEQ ID NO: 51)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTAGCCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE8A4-LC-AA (SEQ IDNO: 52) DIQMTQSPSSLSASVGDRVTITCRASQSIAKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRSPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke8A4 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 164) encoded by thenucleic acid sequence shown in SEQ ID NO: 163.

>KE8A4-VL-NT (SEQ ID NO: 163)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGTTCCATAAGCGTAGCCCGCAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE8A4-VL-AA (SEQ ID NO: 164)DIQMTQSPSSLSASVGDRVTITCRASQSIAKYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFHKRSPQTFG QGTKVEIK

The Ke86G9 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 54) encoded by the nucleic acid sequence shownin SEQ ID NO: 53.

>KE86G9-LC-NT (SEQ ID NO: 53)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAATGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGGGCCGACCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KE86G9-LC-AA (SEQ IDNO: 54) DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYNASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPTTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ke86G9 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 166) encoded by thenucleic acid sequence shown in SEQ ID NO: 165.

>KE86G9-VL-NT (SEQ ID NO: 165)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAATGCATCCAATTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGGCATCCGCGTGGGCCGACCACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KE86G9-VL-AA (SEQ ID NO: 166)DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYNASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQRHPRGPTTFG QGTKVEIK

The Ka3 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 56) encoded by the nucleic acid sequence shownin SEQ ID NO: 55.

>KA3-LC-NT (SEQ ID NO: 55)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCACCCGCGCGCCCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3-LC-AA (SEQ ID NO:56) DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 168) encoded by thenucleic acid sequence shown in SEQ ID NO: 167.

>KA3-VL-NT (SEQ ID NO: 167)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCACCCGCGCGCCCCGAAGACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3-VL-AA (SEQ ID NO: 168)DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ka3A2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 58) encoded by the nucleic acid sequence shownin SEQ ID NO: 57.

>KA3A2-LC-NT (SEQ ID NO: 57)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3A2-LC-AA (SEQ IDNO: 58) DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3A2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 170) encoded by thenucleic acid sequence shown in SEQ ID NO: 169.

>KA3A2-VL-NT (SEQ ID NO: 169)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3A2-VL-AA (SEQ ID NO: 170)DIQMTQSPSSLSASVGDRVTITCRASQSISKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFG QGTKVEIK

The Ka3H3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 60) encoded by the nucleic acid sequence shownin SEQ ID NO: 59.

>KA3H3-LC-NT (SEQ ID NO: 59)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTGCTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGCTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3H3-LC-AA (SEQ IDNO: 60) DIQMTQSPSSLSASVGDRVTITCQASQDIAKYLNWYQQKPGKAPKLLIYAASALQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3H3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 172) encoded by thenucleic acid sequence shown in SEQ ID NO: 171.

>KA3H3-VL-NT (SEQ ID NO: 171)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCGAGTCAGGACATTGCTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGCTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3H3-VL-AA (SEQ ID NO: 172)DIQMTQSPSSLSASVGDRVTITCQASQDIAKYLNWYQQKPGKAPKLLIYAASALQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFG QGTKVEIK

The Ka3A3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 62) encoded by the nucleic acid sequence shownin SEQ ID NO: 61.

>KA3A3-LC-NT (SEQ ID NO: 61)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3A3-LC-AA (SEQ IDNO: 62) DIQMTQSPSSLSASVGDRVTITCRASQSIASYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3A3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 174) encoded by thenucleic acid sequence shown in SEQ ID NO: 173.

>KA3A3-VL-NT (SEQ ID NO: 173)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCTAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3A3-VL-AA (SEQ ID NO: 174)DIQMTQSPSSLSASVGDRVTITCRASQSIASYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ka3H8 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 64) encoded by the nucleic acid sequence shownin SEQ TD NO: 63.

>KA3H8-LC-NT (SEQ ID NO: 63)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCGAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3H8-LC-AA (SEQ IDNO: 64) DIQMTQSPSSLSASVGDRVTITCRASQSIASYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3H8 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 176) encoded by thenucleic acid sequence shown in SEQ ID NO: 175.

>KA3H8-VL-NT (SEQ ID NO: 175)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGCGAGTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCGGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCTCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3H8-VL-AA (SEQ ID NO: 176)DIQMTQSPSSLSASVGDRVTITCRASQSIASYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRSPKTFG QGTKVEIK

The Ka3B2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 66) encoded by the nucleic acid sequence shownin SEQ ID NO: 65.

>KA3B2-LC-NT (SEQ ID NO: 65)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAGTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3B2-LC-AA (SEQ IDNO: 66) DIQMTQSPSSLSASVGDRVTITCRASQNIGKYLNWYQQKPGKAPKLLIYSASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3B2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 178) encoded by thenucleic acid sequence shown in SEQ ID NO: 177.

>KA3B2-VL-NT (SEQ ID NO: 177)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAGTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3B2-VL-AA (SEQ ID NO: 178)DIQMTQSPSSLSASVGDRVTITCRASQNIGKYLNWYQQKPGKAPKLLIYSASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ka3C5 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 68) encoded by the nucleic acid sequence shownin SEQ ID NO: 67.

>KA3C5-LC-NT (SEQ ID NO: 67)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCTCTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCCCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3C5-LC-AA (SEQ IDNO: 68) DIQMTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYSASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3C5 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 180) encoded by thenucleic acid sequence shown in SEQ ID NO: 179.

>KA3C5-VL-NT (SEQ ID NO: 179)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGTAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCTCTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCCCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3C5-VL-AA (SEQ ID NO: 180)DIQMTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYSASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Ka3G2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 70) encoded by the nucleic acid sequence shownin SEQ ID NO: 69.

>KA3G2-LC-NT (SEQ ID NO: 69)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGGGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3G2-LC-AA (SEQ IDNO: 70) DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3G2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 182) encoded by thenucleic acid sequence shown in SEQ ID NO: 181.

>KA3G2-VL-NT (SEQ ID NO: 181)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGATAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGGGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3G2-VL-AA (SEQ ID NO: 182)DIQMTQSPSSLSASVGDRVTITCRASQSIDKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRGPKTFG QGTKVEIK

The Ka3D3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a kappalight chain (SEQ ID NO: 72) encoded by the nucleic acid sequence shownin SEQ ID NO: 71.

>KA3D3-LC-NT (SEQ ID NO: 71)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAA >KA3D3-LC-AA (SEQ IDNO: 72) DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

The Ka3D3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a kappa variable light domain (SEQ ID NO: 184) encoded by thenucleic acid sequence shown in SEQ ID NO: 183.

>KA3D3-VL-NT (SEQ ID NO: 183)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGTAAGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGATGCATCCTCGCGCGCCGAAAACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >KA3D3-VL-AA (SEQ ID NO: 184)DIQMTQSPSSLSASVGDRVTITCRASQSIGKYLNWYQQKPGKAPKLLIYAASRLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQMHPRAPKTFG QGTKVEIK

The Kc4 antibody includes a common heavy chain (SEQ TD NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 74) encoded by the nucleic acid sequence shownin SEQ ID NO: 73.

>KC4-LC-NT (SEQ ID NO: 73)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGTGGTTATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGGTCAGTAATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4-LC-AA (SEQID NO: 74) QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYEVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 186) encoded by thenucleic acid sequence shown in SEQ ID NO: 185.

>KC4-VL-NT (SEQ ID NO: 185)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGTGGTTATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGGTCAGTAATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4-VL-AA (SEQ ID NO: 186)QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYEVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4G11 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 76) encoded by the nucleic acid sequence shownin SEQ TD NO: 75.

>KC4G11-LC-NT (SEQ ID NO: 75)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGGAAGGCGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGGATAGTGATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4G11-LC-AA(SEQ ID NO: 76) QSALTQPASVSGSPGQSITISCTGTSSDVGKANYVSWYQQHPGKAPKLMIYKDSDRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4G11 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ TD NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 188) encoded by thenucleic acid sequence shown in SEQ ID NO: 187.

>KC4G11-VL-NT (SEQ ID NO: 187)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGGGAAGGCGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGGATAGTGATCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4G11-VL-AA (SEQ ID NO: 188)QSALTQPASVSGSPGQSITISCTGTSSDVGKANYVSWYQQHPGKAPKLMIYKDSDRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4C11 antibody includes a common heavy chain (SEQ TD NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 78) encoded by the nucleic acid sequence shownin SEQ TD NO: 77.

>KC4C11-LC-NT (SEQ ID NO: 77)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGGGAATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAATAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4C11-LC-AA(SEQ ID NO: 78) QSALTQPASVSGSPGQSITISCTGTSSDVRGNNYVSWYQQHPGKAPKLMIYENSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4C11 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 190) encoded by thenucleic acid sequence shown in SEQ ID NO: 189.

>KC4C11-VL-NT (SEQ ID NO: 189)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGGGAATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAATAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4C11-VL-AA (SEQ ID NO: 190)QSALTQPASVSGSPGQSITISCTGTSSDVRGNNYVSWYQQHPGKAPKLMIYENSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4A1 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 80) encoded by the nucleic acid sequence shownin SEQ ID NO: 79.

>KC4A1-LC-NT (SEQ ID NO: 79)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGTGCGAGGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4A1-LC-AA(SEQ ID NO: 80) QSALTQPASVSGSPGQSITISCTGTSSDVSARNYVSWYQQHPGKAPKLMIYESSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4A1 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 192) encoded by thenucleic acid sequence shown in SEQ ID NO: 191.

>KC4A1-VL-NT (SEQ ID NO: 191)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGTGCGAGGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4A1-VL-AA (SEQ ID NO: 192)QSALTQPASVSGSPGQSITISCTGTSSDVSARNYVSWYQQHPGKAPKLMIYESSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4A4 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 82) encoded by the nucleic acid sequence shownin SEQ ID NO: 81.

>KC4A4-LC-NT (SEQ ID NO: 81)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTAGAACCAGCAGTGACGTTAATAATACTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGACTAGTGGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4A4-LC-AA(SEQ ID NO: 82) QSALTQPASVSGSPGQSITISCTRTSSDVNNTNYVSWYQQHPGKAPKLMIYKTSGRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4A4 antibody includes a common variable heavy domain (SEQ TD NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 194) encoded by thenucleic acid sequence shown in SEQ ID NO: 193.

>KC4A4-VL-NT (SEQ ID NO: 193)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTAGAACCAGCAGTGACGTTAATAATACTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGACTAGTGGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4A4-VL-AA (SEQ ID NO: 194)QSALTQPASVSGSPGQSITISCTRTSSDVNNTNYVSWYQQHPGKAPKLMIYKTSGRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4E10 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 84) encoded by the nucleic acid sequence shownin SEQ TD NO: 83.

>KC4E10-LC-NT (SEQ ID NO: 83)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAATTCTGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAGTAGTAGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4E10-LC-AA(SEQ ID NO: 84) QSALTQPASVSGSPGQSITISCTGTSSDVNSANYVSWYQQHPGKAPKLMIYKSSSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4E10 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ TD NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 196) encoded by thenucleic acid sequence shown in SEQ ID NO: 195.

>KC4E10-VL-NT (SEQ ID NO: 195)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAATTCTGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAGTAGTAGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4E10-VL-AA (SEQ ID NO: 196)QSALTQPASVSGSPGQSITISCTGTSSDVNSANYVSWYQQHPGKAPKLMIYKSSSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4G9 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 86) encoded by the nucleic acid sequence shownin SEQ TD NO: 85.

>KC4G9-LC-NT (SEQ ID NO: 85)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCCGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGAGAGGAAGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAATAGTACTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4G9-LC-AA(SEQ ID NO: 86) QSALTQPASVSGSPGQSITISCTGTSSDVERKNYVSWYQQHPGKAPKLMIYKNSTRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4G9 antibody includes a common variable heavy domain (SEQ TD NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 198) encoded by thenucleic acid sequence shown in SEQ ID NO: 197.

>KC4G9-VL-NT (SEQ ID NO: 197)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCCGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTGAGAGGAAGAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAATAGTACTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4G9-VL-AA (SEQ ID NO: 198)QSALTQPASVSGSPGQSITISCTGTSSDVERKNYVSWYQQHPGKAPKLMIYKNSTRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4C3 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 88) encoded by the nucleic acid sequence shownin SEQ ID NO: 87.

>KC4C3-LC-NT (SEQ ID NO: 87)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGCGGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAATAGTACTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4C3-LC-AA(SEQ ID NO: 88) QSALTQPASVSGSPGQSITISCTGTSSDVRAANYVSWYQQHPGKAPKLMIYKNSTRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4C3 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 200) encoded by thenucleic acid sequence shown in SEQ ID NO: 199.

>KC4C3-VL-NT (SEQ ID NO: 199)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGCGGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATAAGAATAGTACTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4C3-VL-AA (SEQ ID NO: 200)QSALTQPASVSGSPGQSITISCTGTSSDVRAANYVSWYQQHPGKAPKLMIYKNSTRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4F4 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 90) encoded by the nucleic acid sequence shownin SEQ TD NO: 89.

>KC4F4-LC-NT (SEQ ID NO: 89)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGAGGGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATCAGGATAGTAGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4F4-LC-AA(SEQ ID NO: 90) QSALTQPASVSGSPGQSITISCTGTSSDVRRANYVSWYQQHPGKAPKLMIYQDSSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4F4 antibody includes a common variable heavy domain (SEQ TD NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 202) encoded by thenucleic acid sequence shown in SEQ ID NO: 201.

>KC4F4-VL-NT (SEQ ID NO: 201)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGAGGGCTAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATCAGGATAGTAGTCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4F4-VL-AA (SEQ ID NO: 202)QSALTQPASVSGSPGQSITISCTGTSSDVRRANYVSWYQQHPGKAPKLMIYQDSSRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4B1 antibody includes a common heavy chain (SEQ TD NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 92) encoded by the nucleic acid sequence shownin SEQ TD NO: 91.

>KC4B1-LC-NT (SEQ ID NO: 91)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGCTAATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTGCGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4B1-LC-AA(SEQ ID NO: 92) QSALTQPASVSGSPGQSITISCTGTSSDVRANNYVSWYQQHPGKAPKLMIYESSARPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4B1 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 204) encoded by thenucleic acid sequence shown in SEQ ID NO: 203.

>KC4B1-VL-NT (SEQ ID NO: 203)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTAGGGCTAATAACTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTGCGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4B1-VL-AA (SEQ ID NO: 204)QSALTQPASVSGSPGQSITISCTGTSSDVRANNYVSWYQQHPGKAPKLMIYESSARPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

The Kc4E2 antibody includes a common heavy chain (SEQ ID NO: 2) encodedby the nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 94) encoded by the nucleic acid sequence shownin SEQ ID NO: 93.

>KC4E2-LC-NT (SEQ ID NO: 93)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTTATTATAATAAGTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTC ATAA >KC4E2-LC-AA(SEQ ID NO: 94) QSALTQPASVSGSPGQSITISCTGTSSDVYYNKYVSWYQQHPGKAPKLMIYESSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPKVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQV THEGSTVEKTVAPTECS

The Kc4E2 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 206) encoded by thenucleic acid sequence shown in SEQ ID NO: 205.

>KC4E2-VL-NT (SEQ ID NO: 205)CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGGAACCAGCAGTGACGTTTATTATAATAAGTATGTCTCCTGGTACCAACAGCACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGAGTAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATGATTGGTGGTTCCGCCCCAAGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >KC4E2-VL-AA (SEQ ID NO: 206)QSALTQPASVSGSPGQSITISCTGTSSDVYYNKYVSWYQQHPGKAPKLMIYESSKRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDWWFRPK VFGGGTKLTVL

Anti-CD19 Antibodies

The C2 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 96) encoded by the nucleic acid sequence shownin SEQ ID NO: 95.

>C2-LC-NT (SEQ ID NO: 95)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGAAGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACCAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >C2-LC-AA(SEQ ID NO: 96) NFMLTQPHSVSESPGKTVTISCTRSSGSIEDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDQSLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The C2 antibody includes a common variable heavy domain (SEQ ID NO: 114)encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

>C2-VL-NT (SEQ ID NO: 207)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGAAGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACCAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >C2-VL-AA (SEQ ID NO: 208)NFMLTQPHSVSESPGKTVTISCTRSSGSIEDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDQSLYG WVFGGGTKLTVL

The A6 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 98) encoded by the nucleic acid sequence shownin SEQ ID NO: 97.

>A6-LC-NT (SEQ ID NO: 97)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGGTGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACGTACGACGAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >A6-LC-AA(SEQ ID NO: 98) NFMLTQPHSVSESPGKTVTISCTRSSGSIGDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDESLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The A6 antibody includes a common variable heavy domain (SEQ ID NO: 114)encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 210) encoded by thenucleic acid sequence shown in SEQ ID NO: 209.

>A6-VL-NT (SEQ ID NO: 209)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGGTGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACGTACGACGAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >A6-VL-AA (SEQ ID NO: 210)NFMLTQPHSVSESPGKTVTISCTRSSGSIGDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDESLYG WVFGGGTKLTVL

The C6 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 100) encoded by the nucleic acid sequence shownin SEQ ID NO: 99.

>C6-LC-NT (SEQ ID NO: 99)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCAATGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTTTGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACACCAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >C6-LC-AA(SEQ ID NO: 100) NFMLTQPHSVSESPGKTVTISCTRSSGSINDKYVQWYQQRPGSSPTIVIYFDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDTSLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The C6 antibody includes a common variable heavy domain (SEQ ID NO: 114)encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 212) encoded by thenucleic acid sequence shown in SEQ ID NO: 211.

>C6-VL-NT (SEQ ID NO: 211)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCAATGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTTTGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACACCAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >C6-VL-AA (SEQ ID NO: 212)NFMLTQPHSVSESPGKTVTISCTRSSGSINDKYVQWYQQRPGSSPTIVIYFDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDTSLYG WVFGGGTKLTVL

The C9 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 102) encoded by the nucleic acid sequence shownin SEQ ID NO: 101.

>C9-LC-NT (SEQ ID NO: 101)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGCTGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACGAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >C9-LC-AA(SEQ ID NO: 102) NFMLTQPHSVSESPGKTVTISCTRSSGSIADKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDESLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The C9 antibody includes a common variable heavy domain (SEQ ID NO: 114)encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 214) encoded by thenucleic acid sequence shown in SEQ ID NO: 213.

>C9-VL-NT (SEQ ID NO: 213)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGCTGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACGAGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >C9-VL-AA (SEQ ID NO: 214)NFMLTQPHSVSESPGKTVTISCTRSSGSIADKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDESLYG WVFGGGTKLTVL

The B11 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 104) encoded by the nucleic acid sequence shownin SEQ ID NO: 103.

>B11-LC-NT (SEQ ID NO: 103)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGAAGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACAACAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >B11-LC-AA(SEQ ID NO: 104) NFMLTQPHSVSESPGKTVTISCTRSSGSIEDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDNSLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The B11 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 216) encoded by thenucleic acid sequence shown in SEQ ID NO: 215.

>B11-VL-NT (SEQ ID NO: 215)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGAAGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCATTGTGATCTATTATGATAACGAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGACCTACGACAACAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >B11-VL-AA (SEQ ID NO: 216)NFMLTQPHSVSESPGKTVTISCTRSSGSIEDKYVQWYQQRPGSSPTIVIYYDNERPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQTYDNSLYG WVFGGGTKLTVL

The D11 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 106) encoded by the nucleic acid sequence shownin SEQ ID NO: 105.

>D11-LC-NT (SEQ ID NO: 105)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCAGCATCGATGATAAGTTTGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCACTGTGATCTATTATGATAACATTAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGTCCTATGACGCGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >D11-LC-AA(SEQ ID NO: 106) NFMLTQPHSVSESPGKTVTISCTRSSGSIDDKFVQWYQQRPGSSPTTVIYYDNIRPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDASLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The D11 antibody includes a common variable heavy domain (SEQ ID NO:114) encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

>D11-VL-NT (SEQ ID NO: 217)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCAGCATCGATGATAAGTTTGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCACTGTGATCTATTATGATAACATTAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGTCCTATGACGCGAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >D11-VL-AA (SEQ ID NO: 218)NFMLTQPHSVSESPGKTVTISCTRSSGSIDDKFVQWYQQRPGSSPTTVIYYDNIRPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDASLYG WVFGGGTKLTVL

The B7 antibody includes a common heavy chain (SEQ ID NO: 2) encoded bythe nucleic acid sequence shown in SEQ ID NO: 1 and includes a lambdalight chain (SEQ ID NO: 108) encoded by the nucleic acid sequence shownin SEQ ID NO: 107.

>B7-LC-NT (SEQ ID NO: 107)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGCGGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCACTGTGATCTATGAGGATAACCAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGTCCTATGACAGCAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATG TTCATAA >B7-LC-AA(SEQ ID NO: 108) NFMLTQPHSVSESPGKTVTISCTRSSGSIADKYVQWYQQRPGSSPTTVIYEDNQRPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDSSLYGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS

The B7 antibody includes a common variable heavy domain (SEQ TD NO: 114)encoded by the nucleic acid sequence shown in SEQ ID NO: 113 andincludes a lambda variable light domain (SEQ ID NO: 220) encoded by thenucleic acid sequence shown in SEQ ID NO: 219.

>B7-VL-NT (SEQ ID NO: 219)AATTTTATGCTGACTCAGCCCCACTCTGTGTCGGAGTCTCCGGGGAAGACGGTAACCATCTCCTGCACCCGCAGCAGTGGCTCTATCGCGGATAAGTATGTGCAGTGGTACCAGCAGCGCCCGGGCAGTTCCCCCACCACTGTGATCTATGAGGATAACCAAAGACCCTCTGGGGTCCCTGATCGGTTCTCTGGCTCCATCGACAGCTCCTCCAACTCTGCCTCCCTCACCATCTCTGGACTGAAGACTGAGGACGAGGCTGACTACTACTGTCAGTCCTATGACAGCAGCCTGTATGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >B7-VL-AA (SEQ ID NO: 220)NFMLTQPHSVSESPGKTVTISCTRSSGSIADKYVQWYQQRPGSSPTTVIYEDNQRPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDSSLYG WVFGGGTKLTVL

Dummy Light Chains

The Dummy light chain 1 (SEQ ID NO: 110) is encoded by the nucleic acidsequence shown in SEQ ID NO: 109.

>DUMMY-LC1-NT (SEQ ID NO: 109)CAGTCTGTGTTGACGCAGCCGCCCTCAGTGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAAGCAGCTCCAATATTGAGACTGGTTCTGTATCCTGGTACCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATTTATGACAATAATAAGCGACCCTCAGGGATTCCTGACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGCCGATTATTACTGCGGAACATGGGATGACAGCCTGCCTGGATGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCTTGGAAAGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCATA A >DUMMY-LC1-AA (SEQID NO: 110) QSVLTQPPSVSAAPGQKVTISCSGSSSNIETGSVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDDSLPGWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVT HEGSTVEKTVAPTECS

The Dummy variable light domain 1 (SEQ ID NO: 222) is encoded by thenucleic acid sequence shown in SEQ ID NO: 221.

>DUMMY-VL1-NT (SEQ ID NO: 221)CAGTCTGTGTTGACGCAGCCGCCCTCAGTGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAAGCAGCTCCAATATTGAGACTGGTTCTGTATCCTGGTACCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATTTATGACAATAATAAGCGACCCTCAGGGATTCCTGACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGCCGATTATTACTGCGGAACATGGGATGACAGCCTGCCTGGATGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA >DUMMY-VL1-AA (SEQ ID NO: 222)QSVLTQPPSVSAAPGQKVTISCSGSSSNIETGSVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDDSLPGWV FGGGTKLTVL

The Dummy light chain 2 (SEQ ID NO: 112) is encoded by the nucleic acidsequence shown in SEQ ID NO: 111.

>DUMMY-LC2-NT (SEQ ID NO: 111)GAAATAGTGATGACGCAGTCTCCAGCCACCCTGTCTGTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACGGTTAAGAATAATTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCACCAGGGCCACTGGTATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGTCTGAAGATTTTGCAGTTTATTACTGTCAGCAGTATAACAACTGGTTGCCCATCAACCCCTATACCTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA GTGTTAA >DUMMY-LC2-AA(SEQ ID NO: 112) EIVMTQSPATLSVSPGERATLSCRASQTVKNNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWLPINPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC

The Dummy variable light domain 2 (SEQ ID NO: 224) is encoded by thenucleic acid sequence shown in SEQ ID NO: 223.

>DUMMY-VL2-NT (SEQ ID NO: 223)GAAATAGTGATGACGCAGTCTCCAGCCACCCTGTCTGTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGACGGTTAAGAATAATTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCACCAGGGCCACTGGTATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGTCTGAAGATTTTGCAGTTTATTACTGTCAGCAGTATAACAACTGGTTGCCCATCAACCCCTATACCTTCGGCCAAGGGACCAAGGTGGAAATCAAA >DUMMY-VL2-AA (SEQ ID NO: 224)EIVMTQSPATLSVSPGERATLSCRASQTVKNNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWLPINPY TFGQGTKVEIK

Monovalent Antibodies

In some embodiments, the monovalent antibody 5A3 includes a common heavychain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQID NO: 1, a kappa light chain (SEQ ID NO: 4) encoded by the nucleic acidsequence shown in SEQ ID NO: 3 and a lambda dummy light chain 1 (SEQ IDNO: 110) encoded by the nucleic acid sequence shown in SEQ ID NO: 109.In some embodiments, the monovalent antibody 5A3 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 116) encoded by the nucleic acid sequence shown in SEQ ID NO: 115and a lambda dummy variable light domain 1 (SEQ ID NO: 222) encoded bythe nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody 5A3-M3 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 8) encoded by the nucleicacid sequence shown in SEQ ID NO: 7 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody 5A3-M3 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 120) encoded by the nucleic acid sequence shown in SEQ IDNO: 119 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody 5A3-M5 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 10) encoded by the nucleicacid sequence shown in SEQ ID NO: 9 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody 5A3-M5 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 122) encoded by the nucleic acid sequence shown in SEQ IDNO: 121 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8 includes a common heavychain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQID NO: 1, a kappa light chain (SEQ ID NO: 12) encoded by the nucleicacid sequence shown in SEQ ID NO: 11 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 124) encoded by the nucleic acid sequence shown in SEQ IDNO: 123 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8A2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 18) encoded by the nucleicacid sequence shown in SEQ ID NO: 17 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8A2 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 130) encoded by the nucleic acid sequence shown in SEQ IDNO: 129 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8B2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 16) encoded by the nucleicacid sequence shown in SEQ ID NO: 15 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8B2 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 128) encoded by the nucleic acid sequence shown in SEQ IDNO: 127 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8G11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 46) encoded by the nucleicacid sequence shown in SEQ ID NO: 45 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8G11 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 158) encoded by the nucleic acid sequence shown in SEQ IDNO: 157 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8C4 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 42) encoded by the nucleicacid sequence shown in SEQ ID NO: 41 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8C4 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 154) encoded by the nucleic acid sequence shown in SEQ IDNO: 153 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ke8A3 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 26) encoded by the nucleicacid sequence shown in SEQ ID NO: 25 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ke8A3 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 138) encoded by the nucleic acid sequence shown in SEQ IDNO: 137 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ka3 includes a common heavychain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQID NO: 1, a kappa light chain (SEQ ID NO: 56) encoded by the nucleicacid sequence shown in SEQ ID NO: 55 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ka3 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 168) encoded by the nucleic acid sequence shown in SEQ IDNO: 167 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ka3A3 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 62) encoded by the nucleicacid sequence shown in SEQ ID NO: 61 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ka3A3 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 174) encoded by the nucleic acid sequence shown in SEQ IDNO: 173 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ka3G2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 70) encoded by the nucleicacid sequence shown in SEQ ID NO: 69 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ka3G2 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 182) encoded by the nucleic acid sequence shown in SEQ IDNO: 181 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody Ka3H3 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 60) encoded by the nucleicacid sequence shown in SEQ ID NO: 59 and a lambda dummy light chain 1(SEQ ID NO: 110) encoded by the nucleic acid sequence shown in SEQ IDNO: 109. In some embodiments, the monovalent antibody Ka3H3 includes acommon variable heavy domain (SEQ ID NO: 114) encoded by the nucleicacid sequence shown in SEQ ID NO: 113, a kappa variable light domain(SEQ ID NO: 172) encoded by the nucleic acid sequence shown in SEQ IDNO: 171 and a lambda dummy variable light domain 1 (SEQ ID NO: 222)encoded by the nucleic acid sequence shown in SEQ ID NO: 221.

In some embodiments, the monovalent antibody C2 includes a common heavychain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown in SEQID NO: 1, a kappa dummy light chain 2 (SEQ ID NO: 112) encoded by thenucleic acid sequence shown in SEQ ID NO: 111 and a lambda light chain(SEQ ID NO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO:95. In some embodiments, the monovalent antibody C2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa dummy variable light domain 2(SEQ ID NO: 224) encoded by the nucleic acid sequence shown in SEQ IDNO: 223 and a lambda variable light domain (SEQ ID NO: 208) encoded bythe nucleic acid sequence shown in SEQ ID NO: 207.

Bispecific Antibodies

In some embodiments, the bispecific antibody 5A3xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 4) encoded by the nucleicacid sequence shown in SEQ ID NO: 3 and a lambda light chain (SEQ ID NO:106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105. Insome embodiments, the bispecific antibody 5A3xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 116) encoded by the nucleic acid sequence shown in SEQ ID NO: 115and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody 5A3-M3xD11 includes acommon heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequenceshown in SEQ ID NO: 1, a kappa light chain (SEQ ID NO: 8) encoded by thenucleic acid sequence shown in SEQ ID NO: 7 and a lambda light chain(SEQ ID NO: 106) encoded by the nucleic acid sequence shown in SEQ IDNO: 105. In some embodiments, the bispecific antibody 5A3-M3xD11includes a common variable heavy domain (SEQ ID NO: 114) encoded by thenucleic acid sequence shown in SEQ ID NO: 113, a kappa variable lightdomain (SEQ ID NO: 120) encoded by the nucleic acid sequence shown inSEQ ID NO: 119 and a lambda variable light domain (SEQ ID NO: 218)encoded by the nucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody 5A3-M3xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 8) encoded by the nucleicacid sequence shown in SEQ ID NO: 7 and a lambda light chain (SEQ ID NO:96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. In someembodiments, the bispecific antibody 5A3-M3xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 120) encoded by the nucleic acid sequence shown in SEQ ID NO: 119and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody 5A3-M5xD11 includes acommon heavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequenceshown in SEQ ID NO: 1, a kappa light chain (SEQ ID NO: 10) encoded bythe nucleic acid sequence shown in SEQ ID NO: 9 and a lambda light chain(SEQ ID NO: 106) encoded by the nucleic acid sequence shown in SEQ IDNO: 105. In some embodiments, the bispecific antibody 5A3-M5xD11includes a common variable heavy domain (SEQ ID NO: 114) encoded by thenucleic acid sequence shown in SEQ ID NO: 113, a kappa variable lightdomain (SEQ ID NO: 122) encoded by the nucleic acid sequence shown inSEQ ID NO: 121 and a lambda variable light domain (SEQ ID NO: 218)encoded by the nucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody 5A3-M5xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 10) encoded by the nucleicacid sequence shown in SEQ ID NO: 9 and a lambda light chain (SEQ ID NO:96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. In someembodiments, the bispecific antibody 5A3-M5xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 122) encoded by the nucleic acid sequence shown in SEQ ID NO: 121and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ke8xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 12) encoded by the nucleicacid sequence shown in SEQ ID NO: 11 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 124) encoded by the nucleic acid sequence shown in SEQ ID NO: 123and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 12) encoded by the nucleicacid sequence shown in SEQ ID NO: 11 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 124) encoded by the nucleic acid sequence shown in SEQ ID NO: 123and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8A2xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 18) encoded by the nucleicacid sequence shown in SEQ ID NO: 17 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8A2xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 130) encoded by the nucleic acid sequence shown in SEQ ID NO: 129and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8B2xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 16) encoded by the nucleicacid sequence shown in SEQ ID NO: 15 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8B2xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 128) encoded by the nucleic acid sequence shown in SEQ ID NO: 127and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8G11xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 46) encoded by the nucleicacid sequence shown in SEQ ID NO: 45 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ke8GxC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 158) encoded by the nucleic acid sequence shown in SEQ ID NO: 157and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ke8C4xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 42) encoded by the nucleicacid sequence shown in SEQ ID NO: 41 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8C4xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 154) encoded by the nucleic acid sequence shown in SEQ ID NO: 153and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8C4xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 42) encoded by the nucleicacid sequence shown in SEQ ID NO: 41 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ke8C4xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 154) encoded by the nucleic acid sequence shown in SEQ ID NO: 153and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ke8A3xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 26) encoded by the nucleicacid sequence shown in SEQ ID NO: 25 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ke8A3xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 138) encoded by the nucleic acid sequence shown in SEQ ID NO: 137and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ke8A3xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 26) encoded by the nucleicacid sequence shown in SEQ ID NO: 25 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ke8A3xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 138) encoded by the nucleic acid sequence shown in SEQ ID NO: 137and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ka3xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 56) encoded by the nucleicacid sequence shown in SEQ ID NO: 55 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ka3xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 168) encoded by the nucleic acid sequence shown in SEQ ID NO: 167and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ka3xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 56) encoded by the nucleicacid sequence shown in SEQ ID NO: 55 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ka3xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 168) encoded by the nucleic acid sequence shown in SEQ ID NO: 167and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ka3A3xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 62) encoded by the nucleicacid sequence shown in SEQ ID NO: 61 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ka3A3xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 174) encoded by the nucleic acid sequence shown in SEQ ID NO: 173and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ka3G2xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 70) encoded by the nucleicacid sequence shown in SEQ ID NO: 69 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ka3G2xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 182) encoded by the nucleic acid sequence shown in SEQ ID NO: 181and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ka3G2xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 70) encoded by the nucleicacid sequence shown in SEQ ID NO: 69 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ka3G2xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 182) encoded by the nucleic acid sequence shown in SEQ ID NO: 181and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

In some embodiments, the bispecific antibody Ka3H3xD11 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 60) encoded by the nucleicacid sequence shown in SEQ ID NO: 59 and a lambda light chain (SEQ IDNO: 106) encoded by the nucleic acid sequence shown in SEQ ID NO: 105.In some embodiments, the bispecific antibody Ka3H3xD11 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 172) encoded by the nucleic acid sequence shown in SEQ ID NO: 171and a lambda variable light domain (SEQ ID NO: 218) encoded by thenucleic acid sequence shown in SEQ ID NO: 217.

In some embodiments, the bispecific antibody Ka3H3xC2 includes a commonheavy chain (SEQ ID NO: 2) encoded by the nucleic acid sequence shown inSEQ ID NO: 1, a kappa light chain (SEQ ID NO: 60) encoded by the nucleicacid sequence shown in SEQ ID NO: 59 and a lambda light chain (SEQ IDNO: 96) encoded by the nucleic acid sequence shown in SEQ ID NO: 95. Insome embodiments, the bispecific antibody Ka3H3xC2 includes a commonvariable heavy domain (SEQ ID NO: 114) encoded by the nucleic acidsequence shown in SEQ ID NO: 113, a kappa variable light domain (SEQ IDNO: 172) encoded by the nucleic acid sequence shown in SEQ ID NO: 171and a lambda variable light domain (SEQ ID NO: 208) encoded by thenucleic acid sequence shown in SEQ ID NO: 207.

Definitions

Unless otherwise defined, scientific and technical terms used inconnection with the present invention shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular. Generally,nomenclatures utilized in connection with, and techniques of, cell andtissue culture, molecular biology, and protein and oligo- orpolynucleotide chemistry and hybridization described herein are thosewell-known and commonly used in the art. Standard techniques are usedfor recombinant DNA, oligonucleotide synthesis, and tissue culture andtransformation (e.g., electroporation, lipofection). Enzymatic reactionsand purification techniques are performed according to manufacturer'sspecifications or as commonly accomplished in the art or as describedherein. The foregoing techniques and procedures are generally performedaccording to conventional methods well known in the art and as describedin various general and more specific references that are cited anddiscussed throughout the present specification. See e.g., Sambrook etal. Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y. (1989)). The nomenclaturesutilized in connection with, and the laboratory procedures andtechniques of, analytical chemistry, synthetic organic chemistry, andmedicinal and pharmaceutical chemistry described herein are thosewell-known and commonly used in the art. Standard techniques are usedfor chemical syntheses, chemical analyses, pharmaceutical preparation,formulation, and delivery, and treatment of patients.

As utilized in accordance with the present disclosure, the followingterms, unless otherwise indicated, shall be understood to have thefollowing meanings:

As used herein, the term “antibody” refers to immunoglobulin moleculesand immunologically active portions of immunoglobulin (Ig) molecules,i.e., molecules that contain an antigen binding site that specificallybinds (immunoreacts with) an antigen. By “specifically bind” or“immunoreacts with” or “immunospecifically bind” is meant that theantibody reacts with one or more antigenic determinants of the desiredantigen and does not react with other polypeptides or binds at muchlower affinity (K_(d)>10⁻⁶). Antibodies include, but are not limited to,polyclonal, monoclonal, chimeric, dAb (domain antibody), single chain,F_(ab), F_(ab′) and F_((ab′)2) fragments, scFvs, and an F_(ab)expression library.

The basic antibody structural unit is known to comprise a tetramer. Eachtetramer is composed of two identical pairs of polypeptide chains, eachpair having one “light” (about 25 kDa) and one “heavy” chain (about50-70 kDa). The amino-terminal portion of each chain includes a variableregion of about 100 to 110 or more amino acids primarily responsible forantigen recognition. The carboxy-terminal portion of each chain definesa constant region primarily responsible for effector function. Ingeneral, antibody molecules obtained from humans relate to any of theclasses IgG, IgM, IgA, IgE and IgD, which differ from one another by thenature of the heavy chain present in the molecule. Certain classes havesubclasses as well, such as IgG₁, IgG₂, and others. Furthermore, inhumans, the light chain may be a kappa chain or a lambda chain.

The term “monoclonal antibody” (MAb) or “monoclonal antibodycomposition”, as used herein, refers to a population of antibodymolecules that contain only one molecular species of antibody moleculeconsisting of a unique light chain gene product and a unique heavy chaingene product. In particular, the complementarity determining regions(CDRs) of the monoclonal antibody are identical in all the molecules ofthe population. MAbs contain an antigen binding site capable ofimmunoreacting with a particular epitope of the antigen characterized bya unique binding affinity for it.

The term “antigen-binding site,” or “binding portion” refers to the partof the immunoglobulin molecule that participates in antigen binding. Theantigen binding site is formed by amino acid residues of the N-terminalvariable (“V”) regions of the heavy (“H”) and light (“L”) chains. Threehighly divergent stretches within the V regions of the heavy and lightchains, referred to as “hypervariable regions,” are interposed betweenmore conserved flanking stretches known as “framework regions,” or“FRs”. Thus, the term “FR” refers to amino acid sequences which arenaturally found between, and adjacent to, hypervariable regions inimmunoglobulins. In an antibody molecule, the three hypervariableregions of a light chain and the three hypervariable regions of a heavychain are disposed relative to each other in three dimensional space toform an antigen-binding surface. The antigen-binding surface iscomplementary to the three-dimensional surface of a bound antigen, andthe three hypervariable regions of each of the heavy and light chainsare referred to as “complementarity-determining regions,” or “CDRs.” Theassignment of amino acids to each domain is in accordance with thedefinitions of Kabat Sequences of Proteins of Immunological Interest(National Institutes of Health, Bethesda, Md. (1987 and 1991)), orChothia & Lesk J. Mol. Biol. 196:901-917 (1987), Chothia et al. Nature342:878-883 (1989).

As used herein, the term “epitope” includes any protein determinantcapable of specific binding to an immunoglobulin, an scFv, or a T-cellreceptor. The term “epitope” includes any protein determinant capable ofspecific binding to an immunoglobulin or T-cell receptor. Epitopicdeterminants usually consist of chemically active surface groupings ofmolecules such as amino acids or sugar side chains and usually havespecific three dimensional structural characteristics, as well asspecific charge characteristics. For example, antibodies may be raisedagainst N-terminal or C-terminal peptides of a polypeptide. An antibodyis the to specifically bind an antigen when the dissociation constant is≤1 μM; e.g., ≤100 nM, preferably ≤10 nM and more preferably ≤1 nM.

As used herein, the terms “immunological binding,” and “immunologicalbinding properties” refer to the non-covalent interactions of the typewhich occur between an immunoglobulin molecule and an antigen for whichthe immunoglobulin is specific. The strength, or affinity ofimmunological binding interactions can be expressed in terms of thedissociation constant (K_(d)) of the interaction, wherein a smallerK_(d) represents a greater affinity. Immunological binding properties ofselected polypeptides can be quantified using methods well known in theart. One such method entails measuring the rates of antigen-bindingsite/antigen complex formation and dissociation, wherein those ratesdepend on the concentrations of the complex partners, the affinity ofthe interaction, and geometric parameters that equally influence therate in both directions. Thus, both the “on rate constant” (K_(on)) andthe “off rate constant” (K_(off)) can be determined by calculation ofthe concentrations and the actual rates of association and dissociation.(See Nature 361:186-87 (1993)). The ratio of K_(off)/K_(on) enables thecancellation of all parameters not related to affinity, and is equal tothe dissociation constant K_(d). (See, generally, Davies et al. (1990)Annual Rev Biochem 59:439-473). An antibody of the present invention isthe to specifically bind to its target, when the equilibrium bindingconstant (K_(d)) is ≤1 μM, e.g., ≤100 nM, preferably ≤10 nM, and morepreferably ≤1 nM, as measured by assays such as radioligand bindingassays or similar assays known to those skilled in the art.

The term “isolated polynucleotide” as used herein shall mean apolynucleotide of genomic, cDNA, or synthetic origin or some combinationthereof, which by virtue of its origin the “isolated polynucleotide” (1)is not associated with all or a portion of a polynucleotide in which the“isolated polynucleotide” is found in nature, (2) is operably linked toa polynucleotide which it is not linked to in nature, or (3) does notoccur in nature as part of a larger sequence. Polynucleotides inaccordance with the invention include the nucleic acid moleculesencoding the heavy chain immunoglobulin molecules, and nucleic acidmolecules encoding the light chain immunoglobulin molecules describedherein.

The term “isolated protein” referred to herein means a protein of cDNA,recombinant RNA, or synthetic origin or some combination thereof, whichby virtue of its origin, or source of derivation, the “isolated protein”(1) is not associated with proteins found in nature, (2) is free ofother proteins from the same source, e.g., free of marine proteins, (3)is expressed by a cell from a different species, or (4) does not occurin nature.

The term “polypeptide” is used herein as a generic term to refer tonative protein, fragments, or analogs of a polypeptide sequence. Hence,native protein fragments, and analogs are species of the polypeptidegenus. Polypeptides in accordance with the invention comprise the heavychain immunoglobulin molecules, and the light chain immunoglobulinmolecules described herein, as well as antibody molecules formed bycombinations comprising the heavy chain immunoglobulin molecules withlight chain immunoglobulin molecules, such as kappa light chainimmunoglobulin molecules, and vice versa, as well as fragments andanalogs thereof.

The term “naturally-occurring” as used herein as applied to an objectrefers to the fact that an object can be found in nature. For example, apolypeptide or polynucleotide sequence that is present in an organism(including viruses) that can be isolated from a source in nature andwhich has not been intentionally modified by man in the laboratory orotherwise is naturally-occurring.

The term “operably linked” as used herein refers to positions ofcomponents so described are in a relationship permitting them tofunction in their intended manner. A control sequence “operably linked”to a coding sequence is ligated in such a way that expression of thecoding sequence is achieved under conditions compatible with the controlsequences.

The term “control sequence” as used herein refers to polynucleotidesequences which are necessary to effect the expression and processing ofcoding sequences to which they are ligated. The nature of such controlsequences differs depending upon the host organism in prokaryotes, suchcontrol sequences generally include promoter, ribosomal binding site,and transcription termination sequence in eukaryotes, generally, suchcontrol sequences include promoters and transcription terminationsequence. The term “control sequences” is intended to include, at aminimum, all components whose presence is essential for expression andprocessing, and can also include additional components whose presence isadvantageous, for example, leader sequences and fusion partnersequences. The term “polynucleotide” as referred to herein means apolymeric boron of nucleotides of at least 10 bases in length, eitherribonucleotides or deoxynucleotides or a modified form of either type ofnucleotide. The term includes single and double stranded forms of DNA.

As used herein, the twenty conventional amino acids and theirabbreviations follow conventional usage. See Immunology—A Synthesis (2ndEdition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates,Sunderland Mass. (1991)). Stereoisomers (e.g., D-amino acids) of thetwenty conventional amino acids, unnatural amino acids such as α-,α-disubstituted amino acids, N-alkyl amino acids, lactic acid, and otherunconventional amino acids may also be suitable components forpolypeptides of the present invention. Examples of unconventional aminoacids include: 4 hydroxyproline, γ-carboxyglutamate,ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine,N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine,σ-N-methylarginine, and other similar amino acids and imino acids (e.g.,4-hydroxyproline). In the polypeptide notation used herein, theleft-hand direction is the amino terminal direction and the right-handdirection is the carboxy-terminal direction, in accordance with standardusage and convention.

As applied to polypeptides, the term “substantial identity” means thattwo peptide sequences, when optimally aligned, such as by the programsGAP or BESTFIT using default gap weights, share at least 80 percentsequence identity, preferably at least 90 percent sequence identity,more preferably at least 95 percent sequence identity, and mostpreferably at least 99 percent sequence identity.

Preferably, residue positions which are not identical differ byconservative amino acid substitutions.

Conservative amino acid substitutions refer to the interchangeability ofresidues having similar side chains. For example, a group of amino acidshaving aliphatic side chains is glycine, alanine, valine, leucine, andisoleucine; a group of amino acids having aliphatic-hydroxyl side chainsis serine and threonine; a group of amino acids having amide-containingside chains is asparagine and glutamine; a group of amino acids havingaromatic side chains is phenylalanine, tyrosine, and tryptophan; a groupof amino acids having basic side chains is lysine, arginine, andhistidine; and a group of amino acids having sulfur-containing sidechains is cysteine and methionine. Preferred conservative amino acidssubstitution groups are: valine-leucine-isoleucine,phenylalanine-tyrosine, lysine-arginine, alanine valine,glutamic-aspartic, and asparagine-glutamine.

As discussed herein, minor variations in the amino acid sequences ofantibodies or immunoglobulin molecules are contemplated as beingencompassed by the present invention, providing that the variations inthe amino acid sequence maintain at least 75%, more preferably at least80%, 90%, 95%, and most preferably 99%. In particular, conservativeamino acid replacements are contemplated. Conservative replacements arethose that take place within a family of amino acids that are related intheir side chains. Genetically encoded amino acids are generally dividedinto families: (1) acidic amino acids are aspartate, glutamate; (2)basic amino acids are lysine, arginine, histidine; (3) non-polar aminoacids are alanine, valine, leucine, isoleucine, proline, phenylalanine,methionine, tryptophan, and (4) uncharged polar amino acids are glycine,asparagine, glutamine, cysteine, serine, threonine, tyrosine. Thehydrophilic amino acids include arginine, asparagine, aspartate,glutamine, glutamate, histidine, lysine, serine, and threonine. Thehydrophobic amino acids include alanine, cysteine, isoleucine, leucine,methionine, phenylalanine, proline, tryptophan, tyrosine and valine.Other families of amino acids include (i) serine and threonine, whichare the aliphatic-hydroxy family; (ii) asparagine and glutamine, whichare the amide containing family; (iii) alanine, valine, leucine andisoleucine, which are the aliphatic family; and (iv) phenylalanine,tryptophan, and tyrosine, which are the aromatic family. For example, itis reasonable to expect that an isolated replacement of a leucine withan isoleucine or valine, an aspartate with a glutamate, a threonine witha serine, or a similar replacement of an amino acid with a structurallyrelated amino acid will not have a major effect on the binding orproperties of the resulting molecule, especially if the replacement doesnot involve an amino acid within a framework site. Whether an amino acidchange results in a functional peptide can readily be determined byassaying the specific activity of the polypeptide derivative. Assays aredescribed in detail herein. Fragments or analogs of antibodies orimmunoglobulin molecules can be readily prepared by those of ordinaryskill in the art. Preferred amino- and carboxy-termini of fragments oranalogs occur near boundaries of functional domains. Structural andfunctional domains can be identified by comparison of the nucleotideand/or amino acid sequence data to public or proprietary sequencedatabases. Preferably, computerized comparison methods are used toidentify sequence motifs or predicted protein conformation domains thatoccur in other proteins of known structure and/or function. Methods toidentify protein sequences that fold into a known three-dimensionalstructure are known. Bowie et al. Science 253:164 (1991). Thus, theforegoing examples demonstrate that those of skill in the art canrecognize sequence motifs and structural conformations that may be usedto define structural and functional domains in accordance with theinvention.

Preferred amino acid substitutions are those which: (1) reducesusceptibility to proteolysis, (2) reduce susceptibility to oxidation,(3) alter binding affinity for forming protein complexes, (4) alterbinding affinities, and (4) confer or modify other physicochemical orfunctional properties of such analogs. Analogs can include variousmuteins of a sequence other than the naturally-occurring peptidesequence. For example, single or multiple amino acid substitutions(preferably conservative amino acid substitutions) may be made in thenaturally-occurring sequence (preferably in the portion of thepolypeptide outside the domain(s) forming intermolecular contacts. Aconservative amino acid substitution should not substantially change thestructural characteristics of the parent sequence (e.g., a replacementamino acid should not tend to break a helix that occurs in the parentsequence, or disrupt other types of secondary structure thatcharacterizes the parent sequence). Examples of art-recognizedpolypeptide secondary and tertiary structures are described in Proteins,Structures and Molecular Principles (Creighton, Ed., W. H. Freeman andCompany, New York (1984)); Introduction to Protein Structure (C. Brandenand J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); andThornton et at. Nature 354:105 (1991).

As used herein, the terms “label” or “labeled” refers to incorporationof a detectable marker, e.g., by incorporation of a radiolabeled aminoacid or attachment to a polypeptide of biotinyl moieties that can bedetected by marked avidin (e.g., streptavidin containing a fluorescentmarker or enzymatic activity that can be detected by optical orcalorimetric methods). In certain situations, the label or marker canalso be therapeutic. Various methods of labeling polypeptides andglycoproteins are known in the art and may be used. Examples of labelsfor polypeptides include, but are not limited to, the following:radioisotopes or radionuclides (e.g., ³H, ¹⁴C, ¹⁵N, ³⁵S, ⁹⁰Y, ⁹⁹Tc,¹¹¹In, ¹²⁵I, ¹³¹I), fluorescent labels (e.g., FITC, rhodamine,lanthanide phosphors), enzymatic labels (e.g., horseradish peroxidase,p-galactosidase, luciferase, alkaline phosphatase), chemiluminescent,biotinyl groups, predetermined polypeptide epitopes recognized by asecondary reporter (e.g., leucine zipper pair sequences, binding sitesfor secondary antibodies, metal binding domains, epitope tags). In someembodiments, labels are attached by spacer arms of various lengths toreduce potential steric hindrance. The term “pharmaceutical agent ordrug” as used herein refers to a chemical compound or compositioncapable of inducing a desired therapeutic effect when properlyadministered to a patient.

Other chemistry terms herein are used according to conventional usage inthe art, as exemplified by The McGraw-Hill Dictionary of Chemical Terms(Parker, S., Ed., McGraw-Hill, San Francisco (1985)).

As used herein, “substantially pure” means an object species is thepredominant species present (i.e., on a molar basis it is more abundantthan any other individual species in the composition), and preferably asubstantially purified fraction is a composition wherein the objectspecies comprises at least about 50 percent (on a molar basis) of allmacromolecular species present.

Generally, a substantially pure composition will comprise more thanabout 80 percent of all macromolecular species present in thecomposition, more preferably more than about 85%, 90%, 95%, and 99%.Most preferably, the object species is purified to essential homogeneity(contaminant species cannot be detected in the composition byconventional detection methods) wherein the composition consistsessentially of a single macromolecular species.

The term patient includes human and veterinary subjects.

Antibodies

Various procedures known within the art may be used for the productionof polyclonal or monoclonal antibodies directed against a given target,such as, for example, CD47, a tumor associated antigen or other target,or against derivatives, fragments, analogs homologs or orthologsthereof. (See, for example, Antibodies: A Laboratory Manual, Harlow E,and Lane D, 1988, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y., incorporated herein by reference).

Antibodies are purified by well-known techniques, such as affinitychromatography using protein A or protein G, which provide primarily theIgG fraction of immune serum. Subsequently, or alternatively, thespecific antigen which is the target of the immunoglobulin sought, or anepitope thereof, may be immobilized on a column to purify the immunespecific antibody by immunoaffinity chromatography. Purification ofimmunoglobulins is discussed, for example, by D. Wilkinson (TheScientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14,No. 8 (Apr. 17, 2000), pp. 25-28).

In some embodiments, the antibodies of the invention are monoclonalantibodies. Monoclonal antibodies are generated, for example, by usingthe procedures set forth in the Examples provided herein. Antibodies arealso generated, e.g., by immunizing BALB/c mice with combinations ofcell transfectants expressing high levels of a given target on theirsurface. Hybridomas resulting from myeloma/B cell fusions are thenscreened for reactivity to the selected target.

Monoclonal antibodies are prepared, for example, using hybridomamethods, such as those described by Kohler and Milstein, Nature, 256:495(1975). In a hybridoma method, a mouse, hamster, or other appropriatehost animal, is typically immunized with an immunizing agent to elicitlymphocytes that produce or are capable of producing antibodies thatwill specifically bind to the immunizing agent. Alternatively, thelymphocytes can be immunized in vitro.

The immunizing agent will typically include the protein antigen, afragment thereof or a fusion protein thereof. Generally, eitherperipheral blood lymphocytes are used if cells of human origin aredesired, or spleen cells or lymph node cells are used if non-humanmammalian sources are desired. The lymphocytes are then fused with animmortalized cell line using a suitable fusing agent, such aspolyethylene glycol, to form a hybridoma cell (Goding, MonoclonalAntibodies: Principles and Practice, Academic Press, (1986) pp. 59-103).Immortalized cell lines are usually transformed mammalian cells,particularly myeloma cells of rodent, bovine and human origin. Usually,rat or mouse myeloma cell lines are employed. The hybridoma cells can becultured in a suitable culture medium that preferably contains one ormore substances that inhibit the growth or survival of the unfused,immortalized cells. For example, if the parental cells lack the enzymehypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), theculture medium for the hybridomas typically will include hypoxanthine,aminopterin, and thymidine (“HAT medium”), which substances prevent thegrowth of HGPRT-deficient cells.

Preferred immortalized cell lines are those that fuse efficiently,support stable high level expression of antibody by the selectedantibody-producing cells, and are sensitive to a medium such as HATmedium. More preferred immortalized cell lines are murine myeloma lines,which can be obtained, for instance, from the Salk Institute CellDistribution Center, San Diego, Calif. and the American Type CultureCollection, Manassas, Va. Human myeloma and mouse-human heteromyelomacell lines also have been described for the production of monoclonalantibodies. (See Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al.,Monoclonal Antibody Production Techniques and Applications, MarcelDekker, Inc., New York, (1987) pp. 51-63)).

The culture medium in which the hybridoma cells are cultured can then beassayed for the presence of monoclonal antibodies directed against theantigen. Preferably, the binding specificity of monoclonal antibodiesproduced by the hybridoma cells is determined by immunoprecipitation orby an in vitro binding assay, such as radioimmunoassay (RIA) orenzyme-linked immunoabsorbent assay (ELISA). Such techniques and assaysare known in the art. The binding affinity of the monoclonal antibodycan, for example, be determined by the Scatchard analysis of Munson andPollard, Anal. Biochem., 107:220 (1980). Moreover, in therapeuticapplications of monoclonal antibodies, it is important to identifyantibodies having a high degree of specificity and a high bindingaffinity for the target antigen.

After the desired hybridoma cells are identified, the clones can besubcloned by limiting dilution procedures and grown by standard methods.(See Goding, Monoclonal Antibodies: Principles and Practice, AcademicPress, (1986) pp. 59-103). Suitable culture media for this purposeinclude, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640medium. Alternatively, the hybridoma cells can be grown in vivo asascites in a mammal.

The monoclonal antibodies secreted by the subclones can be isolated orpurified from the culture medium or ascites fluid by conventionalimmunoglobulin purification procedures such as, for example, proteinA-SEPHAROSE® (crosslinked, beaded-form of agarose), hydroxylapatitechromatography, gel electrophoresis, dialysis, or affinitychromatography.

Monoclonal antibodies can also be made by recombinant DNA methods, suchas those described in U.S. Pat. No. 4,816,567. DNA encoding themonoclonal antibodies of the invention can be readily isolated andsequenced using conventional procedures (e.g., by using oligonucleotideprobes that are capable of binding specifically to genes encoding theheavy and light chains of murine antibodies). The hybridoma cells of theinvention serve as a preferred source of such DNA. Once isolated, theDNA can be placed into expression vectors, which are then transfectedinto host cells such as simian COS cells, Chinese hamster ovary (CHO)cells, or myeloma cells that do not otherwise produce immunoglobulinprotein, to obtain the synthesis of monoclonal antibodies in therecombinant host cells. The DNA also can be modified, for example, bysubstituting the coding sequence for human heavy and light chainconstant domains in place of the homologous murine sequences (see U.S.Pat. No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or bycovalently joining to the immunoglobulin coding sequence all or part ofthe coding sequence for a non-immunoglobulin polypeptide. Such anon-immunoglobulin polypeptide can be substituted for the constantdomains of an antibody of the invention, or can be substituted for thevariable domains of one antigen-combining site of an antibody of theinvention to create a chimeric bivalent antibody.

Monoclonal antibodies of the invention include humanized antibodies orhuman antibodies. These antibodies are suitable for administration tohumans without engendering an immune response by the human against theadministered immunoglobulin. Humanized forms of antibodies are chimericimmunoglobulins, immunoglobulin chains or fragments thereof (such as Fv,Fab, Fab′, F(ab′)2 or other antigen-binding subsequences of antibodies)that are principally comprised of the sequence of a humanimmunoglobulin, and contain minimal sequence derived from a non-humanimmunoglobulin. Humanization is performed, e.g., by following the methodof Winter and co-workers (Jones et al., Nature, 321:522-525 (1986);Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science,239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences forthe corresponding sequences of a human antibody. (See also U.S. Pat. No.5,225,539). In some instances, Fv framework residues of the humanimmunoglobulin are replaced by corresponding non-human residues.Humanized antibodies also comprise, e.g., residues which are foundneither in the recipient antibody nor in the imported CDR or frameworksequences. In general, the humanized antibody includes substantially allof at least one, and typically two, variable domains, in which all orsubstantially all of the CDR regions correspond to those of a non-humanimmunoglobulin and all or substantially all of the framework regions arethose of a human immunoglobulin consensus sequence. The humanizedantibody optimally also includes at least a portion of an immunoglobulinconstant region (Fc), typically that of a human immunoglobulin (Jones etal., 1986; Riechmann et al., 1988; and Presta, Curr. Op. Struct. Biol.,2:593-596 (1992)).

Fully human antibodies are antibody molecules in which the entiresequence of both the light chain and the heavy chain, including theCDRs, arise from human genes. Such antibodies are termed “humanantibodies”, or “fully human antibodies” herein. Monoclonal antibodiescan be prepared by using trioma technique; the human B-cell hybridomatechnique (see Kozbor, et al., 1983 Immunol Today 4: 72); and the EBVhybridoma technique to produce monoclonal antibodies (see Cole, et al.,1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc.,pp. 77-96). Monoclonal antibodies may be utilized and may be produced byusing human hybridomas (see Cote, et al., 1983. Proc Natl Acad Sci USA80: 2026-2030) or by transforming human B-cells with Epstein Barr Virusin vitro (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCERTHERAPY, Alan R. Liss, Inc., pp. 77-96).

In addition, human antibodies can also be produced using additionaltechniques, including phage display libraries. (See Hoogenboom andWinter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol.,222:581 (1991)). Similarly, human antibodies can be made by introducinghuman immunoglobulin loci into transgenic animals, e.g., mice in whichthe endogenous immunoglobulin genes have been partially or completelyinactivated. Upon challenge, human antibody production is observed,which closely resembles that seen in humans in all respects, includinggene rearrangement, assembly, and antibody repertoire. This approach isdescribed, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806;5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al.,Bio/Technology 10, 779-783 (1992); Lonberg et al., Nature 368 856-859(1994); Morrison, Nature 368, 812-13 (1994); Fishwild et al, NatureBiotechnology 14, 845-51 (1996); Neuberger, Nature Biotechnology 14, 826(1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13 65-93 (1995).

Human antibodies may additionally be produced using transgenic nonhumananimals which are modified so as to produce fully human antibodiesrather than the animal's endogenous antibodies in response to challengeby an antigen. (See PCT publication WO94/02602). The endogenous genesencoding the heavy and light immunoglobulin chains in the nonhuman hosthave been incapacitated, and active loci encoding human heavy and lightchain immunoglobulins are inserted into the host's genome. The humangenes are incorporated, for example, using yeast artificial chromosomescontaining the requisite human DNA segments. An animal which providesall the desired modifications is then obtained as progeny bycrossbreeding intermediate transgenic animals containing fewer than thefull complement of the modifications. An example of such a nonhumananimal is a mouse termed the Xenomouse™ as disclosed in PCT publicationsWO 96/33735 and WO 96/34096. This animal produces B cells which secretefully human immunoglobulins. The antibodies can be obtained directlyfrom the animal after immunization with an immunogen of interest, as,for example, a preparation of a polyclonal antibody, or alternativelyfrom immortalized B cells derived from the animal, such as hybridomasproducing monoclonal antibodies. Additionally, the genes encoding theimmunoglobulins with human variable regions can be recovered andexpressed to obtain the antibodies directly, or can be further modifiedto obtain analogs of antibodies such as, for example, single chain Fv(scFv) molecules.

An example of a method of producing a nonhuman host, exemplified as amouse, lacking expression of an endogenous immunoglobulin heavy chain isdisclosed in U.S. Pat. No. 5,939,598. It can be obtained by a method,which includes deleting the J segment genes from at least one endogenousheavy chain locus in an embryonic stem cell to prevent rearrangement ofthe locus and to prevent formation of a transcript of a rearrangedimmunoglobulin heavy chain locus, the deletion being effected by atargeting vector containing a gene encoding a selectable marker; andproducing from the embryonic stem cell a transgenic mouse whose somaticand germ cells contain the gene encoding the selectable marker.

One method for producing an antibody of interest, such as a humanantibody, is disclosed in U.S. Pat. No. 5,916,771. This method includesintroducing an expression vector that contains a nucleotide sequenceencoding a heavy chain into one mammalian host cell in culture,introducing an expression vector containing a nucleotide sequenceencoding a light chain into another mammalian host cell, and fusing thetwo cells to form a hybrid cell. The hybrid cell expresses an antibodycontaining the heavy chain and the light chain.

In a further improvement on this procedure, a method for identifying aclinically relevant epitope on an immunogen and a correlative method forselecting an antibody that binds specifically to the relevant epitopewith high affinity are disclosed in PCT publication WO 99/53049.

The antibody can be expressed by a vector containing a DNA segmentencoding the single chain antibody described above.

These can include vectors, liposomes, naked DNA, adjuvant-assisted DNA.gene gun, catheters, etc. Vectors include chemical conjugates such asdescribed in WO 93/64701, which has targeting moiety (e.g., a ligand toa cellular surface receptor), and a nucleic acid binding moiety (e.g.,polylysine), viral vector (e.g., a DNA or RNA viral vector), fusionproteins such as described in PCT/US 95/02140 (WO 95/22618) which is afusion protein containing a target moiety (e.g., an antibody specificfor a target cell) and a nucleic acid binding moiety (e.g., aprotamine), plasmids, phage, etc. The vectors can be chromosomal,non-chromosomal or synthetic.

Preferred vectors include viral vectors, fusion proteins and chemicalconjugates. Retroviral vectors include moloney murine leukemia viruses.DNA viral vectors are preferred. These vectors include pox vectors suchas orthopox or avipox vectors, herpesvirus vectors such as a herpessimplex I virus (HSV) vector (see Geller, A. I. et al., J. Neurochem,64:487 (1995); Lim, F., et al., in DNA Cloning: Mammalian Systems, D.Glover, Ed. (Oxford Univ. Press, Oxford England) (1995); Geller, A. I.et al., Proc Natl. Acad. Sci.: U.S.A. 90:7603 (1993); Geller, A. I., etal., Proc Natl. Acad. Sci USA 87:1149 (1990), Adenovirus Vectors (seeLeGal LaSalle et al., Science, 259:988 (1993); Davidson, et al., Nat.Genet 3:219 (1993); Yang, et al., J. Virol. 69:2004 (1995) andAdeno-associated Virus Vectors (see Kaplitt, M. G. et al., Nat. Genet.8:148 (1994).

Pox viral vectors introduce the gene into the cells cytoplasm. Avipoxvirus vectors result in only a short term expression of the nucleicacid. Adenovirus vectors, adeno-associated virus vectors and herpessimplex virus (HSV) vectors are preferred for introducing the nucleicacid into neural cells. The adenovirus vector results in a shorter termexpression (about 2 months) than adeno-associated virus (about 4months), which in turn is shorter than HSV vectors. The particularvector chosen will depend upon the target cell and the condition beingtreated. The introduction can be by standard techniques, e.g.,infection, transfection, transduction or transformation. Examples ofmodes of gene transfer include e.g., naked DNA, CaPO₄ precipitation,DEAE dextran, electroporation, protoplast fusion, lipofection, cellmicroinjection, and viral vectors.

The vector can be employed to target essentially any desired targetcell. For example, stereotaxic injection can be used to direct thevectors (e.g., adenovirus, HSV) to a desired location. Additionally, theparticles can be delivered by intracerebroventricular (icv) infusionusing a minipump infusion system, such as a SynchroMed Infusion System.A method based on bulk flow, termed convection, has also proveneffective at delivering large molecules to extended areas of the brainand may be useful in delivering the vector to the target cell. (See Boboet al., Proc. Natl. Acad. Sci. USA 91:2076-2080 (1994); Morrison et al.,Am. J. Physiol. 266:292-305 (1994)). Other methods that can be usedinclude catheters, intravenous, parenteral, intraperitoneal andsubcutaneous injection, and oral or other known routes ofadministration.

Bispecific antibodies are antibodies that have binding specificities forat least two different antigens. In the present case, one of the bindingspecificities is for a target such as CD47 or any fragment thereof. Thesecond binding target is any other antigen, and advantageously is acell-surface protein or receptor or receptor subunit.

Methods for making bispecific antibodies are known in the art.Traditionally, the recombinant production of bispecific antibodies isbased on the co-expression of two immunoglobulin heavy-chain/light-chainpairs, where the two heavy chains have different specificities (Milsteinand Cuello, Nature, 305:537-539 (1983)). Because of the randomassortment of immunoglobulin heavy and light chains, these hybridomas(quadromas) produce a potential mixture often different antibodymolecules, of which only one has the correct bispecific structure. Thepurification of the correct molecule is usually accomplished by affinitychromatography steps. Similar procedures are disclosed in WO 93/08829,published 13 May 1993, and in Traunecker et al., EMBO J., 10:3655-3659(1991).

Bispecific and/or monovalent antibodies of the invention can be madeusing any of a variety of art-recognized techniques, including thosedisclosed in co-pending application WO 2012/023053, filed Aug. 16, 2011,the contents of which are hereby incorporated by reference in theirentirety. The methods described in WO 2012/023053 generate bispecificantibodies that are identical in structure to a human immunoglobulin.This type of molecule is composed of two copies of a unique heavy chainpolypeptide, a first light chain variable region fused to a constantKappa domain and second light chain variable region fused to a constantLambda domain. Each combining site displays a different antigenspecificity to which both the heavy and light chain contribute. Thelight chain variable regions can be of the Lambda or Kappa family andare preferably fused to a Lambda and Kappa constant domains,respectively. This is preferred in order to avoid the generation ofnon-natural polypeptide junctions. However it is also possible to obtainbispecific antibodies of the invention by fusing a Kappa light chainvariable domain to a constant Lambda domain for a first specificity andfusing a Lambda light chain variable domain to a constant Kappa domainfor the second specificity. The bispecific antibodies described in WO2012/023053 are referred to as IgGκλ antibodies or “κλ bodies,” a newfully human bispecific IgG format. This κλ-body format allows theaffinity purification of a bispecific antibody that is undistinguishablefrom a standard IgG molecule with characteristics that areundistinguishable from a standard monoclonal antibody and, therefore,favorable as compared to previous formats.

An essential step of the method is the identification of two antibody Fvregions (each composed by a variable light chain and variable heavychain domain) having different antigen specificities that share the sameheavy chain variable domain. Numerous methods have been described forthe generation of monoclonal antibodies and fragments thereof. (See,e.g., Antibodies: A Laboratory Manual, Harlow E, and Lane D, 1988, ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y., incorporatedherein by reference). Fully human antibodies are antibody molecules inwhich the sequence of both the light chain and the heavy chain,including the CDRs 1 and 2, arise from human genes. The CDR3 region canbe of human origin or designed by synthetic means. Such antibodies aretermed “human antibodies”, or “fully human antibodies” herein. Humanmonoclonal antibodies can be prepared by using the trioma technique; thehuman B-cell hybridoma technique (see Kozbor, et al., 1983 Immunol Today4: 72); and the EBV hybridoma technique to produce human monoclonalantibodies (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCERTHERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies maybe utilized and may be produced by using human hybridomas (see Cote, etal., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or by transforminghuman B-cells with Epstein Barr Virus in vitro (see Cole, et al., 1985In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp.77-96).

Monoclonal antibodies are generated, e.g., by immunizing an animal witha target antigen or an immunogenic fragment, derivative or variantthereof. Alternatively, the animal is immunized with cells transfectedwith a vector containing a nucleic acid molecule encoding the targetantigen, such that the target antigen is expressed and associated withthe surface of the transfected cells. A variety of techniques arewell-known in the art for producing xenogenic non-human animals. Forexample, see U.S. Pat. No. 6,075,181 and U.S. Pat. No. 6,150,584, whichis hereby incorporated by reference in its entirety.

Alternatively, the antibodies are obtained by screening a library thatcontains antibody or antigen binding domain sequences for binding to thetarget antigen. This library is prepared, e.g., in bacteriophage asprotein or peptide fusions to a bacteriophage coat protein that isexpressed on the surface of assembled phage particles and the encodingDNA sequences contained within the phage particles (i.e., “phagedisplayed library”).

Hybridomas resulting from myeloma/B cell fusions are then screened forreactivity to the target antigen. Monoclonal antibodies are prepared,for example, using hybridoma methods, such as those described by Kohlerand Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse,hamster, or other appropriate host animal, is typically immunized withan immunizing agent to elicit lymphocytes that produce or are capable ofproducing antibodies that will specifically bind to the immunizingagent. Alternatively, the lymphocytes can be immunized in vitro.

Although not strictly impossible, the serendipitous identification ofdifferent antibodies having the same heavy chain variable domain butdirected against different antigens is highly unlikely. Indeed, in mostcases the heavy chain contributes largely to the antigen binding surfaceand is also the most variable in sequence. In particular the CDR3 on theheavy chain is the most diverse CDR in sequence, length and structure.Thus, two antibodies specific for different antigens will almostinvariably carry different heavy chain variable domains.

The methods disclosed in co-pending application WO 2012/023053 overcomesthis limitation and greatly facilitates the isolation of antibodieshaving the same heavy chain variable domain by the use of antibodylibraries in which the heavy chain variable domain is the same for allthe library members and thus the diversity is confined to the lightchain variable domain. Such libraries are described, for example, inco-pending applications WO 2010/135558 and WO 2011/084255, each of whichis hereby incorporated by reference in its entirety. However, as thelight chain variable domain is expressed in conjunction with the heavyvariable domain, both domains can contribute to antigen binding. Tofurther facilitate the process, antibody libraries containing the sameheavy chain variable domain and either a diversity of Lambda variablelight chains or Kappa variable light chains can be used in parallel forin vitro selection of antibodies against different antigens. Thisapproach enables the identification of two antibodies having a commonheavy chain but one carrying a Lambda light chain variable domain andthe other a Kappa light chain variable domain that can be used asbuilding blocks for the generation of a bispecific antibody in the fullimmunoglobulin format of the invention. The bispecific antibodies of theinvention can be of different Isotypes and their Fc portion can bemodified in order to alter the bind properties to different Fc receptorsand in this way modify the effectors functions of the antibody as wellas it pharmacokinetic properties. Numerous methods for the modificationof the Fc portion have been described and are applicable to antibodiesof the invention. (see for example Strohl, W R Curr Opin Biotechnol 2009(6):685-91; U.S. Pat. No. 6,528,624; PCT/US2009/0191199 filed Jan. 9,2009). The methods of the invention can also be used to generatebispecific antibodies and antibody mixtures in a F(ab′)2 format thatlacks the Fc portion.

The common heavy chain and two different light chains are co-expressedinto a single cell to allow for the assembly of a bispecific antibody ofthe invention. If all the polypeptides get expressed at the same leveland get assembled equally well to form an immunoglobulin molecule thenthe ratio of monospecific (same light chains) and bispecific (twodifferent light chains) should be 50%. However, it is likely thatdifferent light chains are expressed at different levels and/or do notassemble with the same efficiency. Therefore, a means to modulate therelative expression of the different polypeptides is used to compensatefor their intrinsic expression characteristics or different propensitiesto assemble with the common heavy chain. This modulation can be achievedvia promoter strength, the use of internal ribosome entry sites (IRES)featuring different efficiencies or other types of regulatory elementsthat can act at transcriptional or translational levels as well asacting on mRNA stability. Different promoters of different strengthcould include CMV (Immediate-early Cytomegalovirus virus promoter);EF1-1α (Human elongation factor 1α-subunit promoter); Ubc (Humanubiquitin C promoter); SV40 (Simian virus 40 promoter). Different IREShave also been described from mammalian and viral origin. (See e.g.,Hellen C U and Sarnow P. Genes Dev 2001 15: 1593-612). These IRES cangreatly differ in their length and ribosome recruiting efficiency.Furthermore, it is possible to further tune the activity by introducingmultiple copies of an IRES (Stephen et al. 2000 Proc Natl Acad Sci USA97: 1536-1541). The modulation of the expression can also be achieved bymultiple sequential transfections of cells to increase the copy numberof individual genes expressing one or the other light chain and thusmodify their relative expressions. The Examples provided hereindemonstrate that controlling the relative expression of the differentchains is critical for maximizing the assembly and overall yield of thebispecific antibody.

The co-expression of the heavy chain and two light chains generates amixture of three different antibodies into the cell culture supernatant:two monospecific bivalent antibodies and one bispecific bivalentantibody. The latter has to be purified from the mixture to obtain themolecule of interest. The method described herein greatly facilitatesthis purification procedure by the use of affinity chromatography mediathat specifically interact with the Kappa or Lambda light chain constantdomains such as the CAPTURESELECT™ Fab Kappa (Fab-Kappa affinitychromatography media) and CAPTURESELECT™ Fab Lambda (Fab-Lambda affinitychromatography media) affinity matrices (BAC BV, Holland). Thismulti-step affinity chromatography purification approach is efficientand generally applicable to antibodies of the invention. This is insharp contrast to specific purification methods that have to bedeveloped and optimized for each bispecific antibodies derived fromquadromas or other cell lines expressing antibody mixtures. Indeed, ifthe biochemical characteristics of the different antibodies in themixtures are similar, their separation using standard chromatographytechnique such as ion exchange chromatography can be challenging or notpossible at all.

Other suitable purification methods include those disclosed inco-pending application PCT/IB2012/003028, filed on Oct. 19, 2012,published as WO2013/088259, the contents of which are herebyincorporated by reference in their entirety.

In other embodiments of producing bispecific antibodies, antibodyvariable domains with the desired binding specificities(antibody-antigen combining sites) can be fused to immunoglobulinconstant domain sequences. The fusion preferably is with animmunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the firstheavy-chain constant region (CH1) containing the site necessary forlight-chain binding present in at least one of the fusions. DNAsencoding the immunoglobulin heavy-chain fusions and, if desired, theimmunoglobulin light chain, are inserted into separate expressionvectors, and are co-transfected into a suitable host organism. Forfurther details of generating bispecific antibodies see, for example,Suresh et al., Methods in Enzymology, 121:210 (1986).

According to another approach described in WO 96/27011, the interfacebetween a pair of antibody molecules can be engineered to maximize thepercentage of heterodimers which are recovered from recombinant cellculture. The preferred interface includes at least a part of the CH3region of an antibody constant domain. In this method, one or more smallamino acid side chains from the interface of the first antibody moleculeare replaced with larger side chains (e.g., tyrosine or tryptophan).Compensatory “cavities” of identical or similar size to the large sidechain(s) are created on the interface of the second antibody molecule byreplacing large amino acid side chains with smaller ones (e.g., alanineor threonine). This provides a mechanism for increasing the yield of theheterodimer over other unwanted end-products such as homodimers.

Techniques for generating bispecific antibodies from antibody fragmentshave been described in the literature. For example, bispecificantibodies can be prepared using chemical linkage. The bispecificantibodies produced can be used as agents for the selectiveimmobilization of enzymes.

Various techniques for making and isolating bispecific antibodyfragments directly from recombinant cell culture have also beendescribed. For example, bispecific antibodies have been produced usingleucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992).The leucine zipper peptides from the Fos and Jun proteins were linked tothe Fab′ portions of two different antibodies by gene fusion. Theantibody homodimers were reduced at the hinge region to form monomersand then re-oxidized to form the antibody heterodimers. This method canalso be utilized for the production of antibody homodimers. The“diabody” technology described by Hollinger et al., Proc. Natl. Acad.Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism formaking bispecific antibody fragments. The fragments comprise aheavy-chain variable domain (V_(H)) connected to a light-chain variabledomain (V_(L)) by a linker which is too short to allow pairing betweenthe two domains on the same chain. Accordingly, the V_(H) and V_(L)domains of one fragment are forced to pair with the complementary V_(L)and V_(H) domains of another fragment, thereby forming twoantigen-binding sites. Another strategy for making bispecific antibodyfragments by the use of single-chain Fv (sFv) dimers has also beenreported. See, Gruber et al., J. Immunol. 152:5368 (1994).

Antibodies with more than two valencies are contemplated. For example,trispecific antibodies can be prepared. Tutt et al., J. Immunol. 147:60(1991).

Exemplary bispecific antibodies can bind to two different epitopes, atleast one of which originates in the protein antigen of the invention.Alternatively, an anti-antigenic arm of an immunoglobulin molecule canbe combined with an arm which binds to a triggering molecule on aleukocyte such as a T-cell receptor molecule (e.g., CD2, CD3, CD28, orB7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32)and FcγRIII (CD16) so as to focus cellular defense mechanisms to thecell expressing the particular antigen. Bispecific antibodies can alsobe used to direct cytotoxic agents to cells which express a particularantigen. These antibodies possess an antigen-binding arm and an armwhich binds a cytotoxic agent or a radionuclide chelator, such asEOTUBE, DPTA, DOTA, or TETA. Another bispecific antibody of interestbinds the protein antigen described herein and further binds tissuefactor (TF).

Heteroconjugate antibodies are also within the scope of the presentinvention. Heteroconjugate antibodies are composed of two covalentlyjoined antibodies. Such antibodies have, for example, been proposed totarget immune system cells to unwanted cells (see U.S. Pat. No.4,676,980), and for treatment of HIV infection (see WO 91/00360; WO92/200373; EP 03089). It is contemplated that the antibodies can beprepared in vitro using known methods in synthetic protein chemistry,including those involving crosslinking agents. For example, immunotoxinscan be constructed using a disulfide exchange reaction or by forming athioether bond. Examples of suitable reagents for this purpose includeiminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, forexample, in U.S. Pat. No. 4,676,980.

It can be desirable to modify the antibody of the invention with respectto effector function, so as to enhance, e.g., the effectiveness of theantibody in treating cancer and/or other diseases and disordersassociated with aberrant CD47 expression and/or activity. For example,cysteine residue(s) can be introduced into the Fc region, therebyallowing interchain disulfide bond formation in this region. Thehomodimeric antibody thus generated can have improved internalizationcapability and/or increased complement-mediated cell killing andantibody-dependent cellular cytotoxicity (ADCC). (See Caron et al., J.Exp Med., 176: 1191-1195 (1992) and Shopes, J. Immunol., 148: 2918-2922(1992)). Alternatively, an antibody can be engineered that has dual Fcregions and can thereby have enhanced complement lysis and ADCCcapabilities. (See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230(1989)).

The invention also pertains to immunoconjugates comprising an antibodyconjugated to a cytotoxic agent such as a toxin (e.g., an enzymaticallyactive toxin of bacterial, fungal, plant, or animal origin, or fragmentsthereof), or a radioactive isotope (i.e., a radioconjugate).

Enzymatically active toxins and fragments thereof that can be usedinclude diphtheria A chain, nonbinding active fragments of diphtheriatoxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain,abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordiiproteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII,and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonariaofficinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin,enomycin, and the tricothecenes. A variety of radionuclides areavailable for the production of radioconjugated antibodies. Examplesinclude ²¹²Bi, ¹³¹, ¹³¹In, ⁹⁰Y, and ¹⁸⁶Re.

Conjugates of the antibody and cytotoxic agent are made using a varietyof bifunctional protein-coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glutareldehyde), bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin canbe prepared as described in Vitetta et al., Science 238: 1098 (1987).Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. (See WO94/11026).

Those of ordinary skill in the art will recognize that a large varietyof possible moieties can be coupled to the resultant antibodies of theinvention. (See, for example, “Conjugate Vaccines”, Contributions toMicrobiology and Immunology, J. M. Cruse and R. E. Lewis, Jr (eds),Carger Press, New York, (1989), the entire contents of which areincorporated herein by reference).

Coupling may be accomplished by any chemical reaction that will bind thetwo molecules so long as the antibody and the other moiety retain theirrespective activities. This linkage can include many chemicalmechanisms, for instance covalent binding, affinity binding,intercalation, coordinate binding and complexation. The preferredbinding is, however, covalent binding. Covalent binding can be achievedeither by direct condensation of existing side chains or by theincorporation of external bridging molecules. Many bivalent orpolyvalent linking agents are useful in coupling protein molecules, suchas the antibodies of the present invention, to other molecules. Forexample, representative coupling agents can include organic compoundssuch as thioesters, carbodiimides, succinimide esters, diisocyanates,glutaraldehyde, diazobenzenes and hexamethylene diamines. This listingis not intended to be exhaustive of the various classes of couplingagents known in the art but, rather, is exemplary of the more commoncoupling agents. (See Killen and Lindstrom, Jour. Immun. 133:1335-2549(1984); Jansen et al., Immunological Reviews 62:185-216 (1982); andVitetta et al., Science 238:1098 (1987).

Preferred linkers are described in the literature. (See, for example,Ramakrishnan, S. et al., Cancer Res. 44:201-208 (1984) describing use ofMBS (M-maleimidobenzoyl-N-hydroxysuccinimide ester). See also, U.S. Pat.No. 5,030,719, describing use of halogenated acetyl hydrazide derivativecoupled to an antibody by way of an oligopeptide linker. Particularlypreferred linkers include: (i) EDC (1-ethyl-3-(3-dimethylamino-propyl)carbodiimide hydrochloride; (ii) SMPT(4-succinimidyloxycarbonyl-alpha-methyl-alpha-(2-pridyl-dithio)-toluene(Pierce Chem. Co., Cat. (21558G); (iii) SPDP (succinimidyl-6[3-(2-pyridyldithio) propionamido]hexanoate (Pierce Chem. Co., Cat#21651G); (iv) Sulfo-LC-SPDP (sulfosuccinimidyl 6[3-(2-pyridyldithio)-propianamide] hexanoate (Pierce Chem. Co. Cat.#2165-G); and (v) sulfo-NHS (N-hydroxysulfo-succinimide: Pierce Chem.Co., Cat. #24510) conjugated to EDC.

The linkers described above contain components that have differentattributes, thus leading to conjugates with differing physio-chemicalproperties. For example, sulfo-NHS esters of alkyl carboxylates are morestable than sulfo-NHS esters of aromatic carboxylates. NHS-estercontaining linkers are less soluble than sulfo-NHS esters. Further, thelinker SMPT contains a sterically hindered disulfide bond, and can formconjugates with increased stability. Disulfide linkages, are in general,less stable than other linkages because the disulfide linkage is cleavedin vitro, resulting in less conjugate available. Sulfo-NHS, inparticular, can enhance the stability of carbodimide couplings.Carbodimide couplings (such as EDC) when used in conjunction withsulfo-NHS, forms esters that are more resistant to hydrolysis than thecarbodimide coupling reaction alone.

The antibodies disclosed herein can also be formulated asimmunoliposomes. Liposomes containing the antibody are prepared bymethods known in the art, such as described in Epstein et al., Proc.Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl Acad.Sci. USA, 77: 4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545.Liposomes with enhanced circulation time are disclosed in U.S. Pat. No.5,013,556.

Particularly useful liposomes can be generated by the reverse-phaseevaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol, and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter. Fab′ fragments of the antibody of the present invention can beconjugated to the liposomes as described in Martin et al., J. Biol.Chem., 257: 286-288 (1982) via a disulfide-interchange reaction.

Use of Anti-CD47 Antibodies

It will be appreciated that administration of therapeutic entities inaccordance with the invention will be administered with suitablecarriers, excipients, and other agents that are incorporated intoformulations to provide improved transfer, delivery, tolerance, and thelike. A multitude of appropriate formulations can be found in theformulary known to all pharmaceutical chemists: Remington'sPharmaceutical Sciences (15th ed, Mack Publishing Company, Easton, Pa.(1975)), particularly Chapter 87 by Blaug, Seymour, therein. Theseformulations include, for example, powders, pastes, ointments, jellies,waxes, oils, lipids, lipid (cationic or anionic) containing vesicles(such as LIPOFECTIN™ (transfection reagent), DNA conjugates, anhydrousabsorption pastes, oil-in-water and water-in-oil emulsions, emulsionscarbowax (polyethylene glycols of various molecular weights), semi-solidgels, and semisolid mixtures containing carbo wax. Any of the foregoingmixtures may be appropriate in treatments and therapies in accordancewith the present invention, provided that the active ingredient in theformulation is not inactivated by the formulation and the formulation isphysiologically compatible and tolerable with the route ofadministration. See also Baldrick P. “Pharmaceutical excipientdevelopment: the need for preclinical guidance.” Regul. ToxicolPharmacol. 32(2):210-8 (2000), Wang W. “Lyophilization and developmentof solid protein pharmaceuticals.” Int. J. Pharm. 203(1-2):1-60 (2000),Charman W N “Lipids, lipophilic drugs, and oral drug delivery-someemerging concepts.” J Pharm Sci. 89(8):967-78 (2000), Powell et al.“Compendium of excipients for parenteral formulations” PDA J Pharm SciTechnol. 52:238-311 (1998) and the citations therein for additionalinformation related to formulations, excipients and carriers well knownto pharmaceutical chemists.

Therapeutic formulations of the invention, which include an antibody ofthe invention, are used to treat or alleviate a symptom associated witha cancer, such as, by way of non-limiting example, leukemias, lymphomas,breast cancer, colon cancer, ovarian cancer, bladder cancer, prostatecancer, glioma, lung & bronchial cancer, colorectal cancer, pancreaticcancer, esophageal cancer, liver cancer, urinary bladder cancer, kidneyand renal pelvis cancer, oral cavity & pharynx cancer, uterine corpuscancer, and/or melanoma The present invention also provides methods oftreating or alleviating a symptom associated with a cancer. Atherapeutic regimen is carried out by identifying a subject, e.g., ahuman patient suffering from (or at risk of developing) a cancer, usingstandard methods.

Efficaciousness of treatment is determined in association with any knownmethod for diagnosing or treating the particular immune-relateddisorder. Alleviation of one or more symptoms of the immune-relateddisorder indicates that the antibody confers a clinical benefit.

Methods for the screening of antibodies that possess the desiredspecificity include, but are not limited to, enzyme linked immunosorbentassay (ELISA) and other immunologically mediated techniques known withinthe art.

Antibodies directed against a target such as CD47, a tumor associatedantigen or other antigen (or a fragment thereof) may be used in methodsknown within the art relating to the localization and/or quantitation ofthese targets, e.g., for use in measuring levels of these targets withinappropriate physiological samples, for use in diagnostic methods, foruse in imaging the protein, and the like). In a given embodiment,antibodies specific any of these targets, or derivative, fragment,analog or homolog thereof, that contain the antibody derived antigenbinding domain, are utilized as pharmacologically active compounds(referred to hereinafter as “Therapeutics”).

An antibody of the invention can be used to isolate a particular targetusing standard techniques, such as immunoaffinity, chromatography orimmunoprecipitation. Antibodies of the invention (or a fragment thereof)can be used diagnostically to monitor protein levels in tissue as partof a clinical testing procedure, e.g., to determine the efficacy of agiven treatment regimen. Detection can be facilitated by coupling (i.e.,physically linking) the antibody to a detectable substance. Examples ofdetectable substances include various enzymes, prosthetic groups,fluorescent materials, luminescent materials, bioluminescent materials,and radioactive materials. Examples of suitable enzymes includehorseradish peroxidase, alkaline phosphatase, 3-galactosidase, oracetylcholinesterase; examples of suitable prosthetic group complexesinclude streptavidin/biotin and avidin/biotin; examples of suitablefluorescent materials include umbelliferone, fluorescein, fluoresceinisothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; an example of a luminescent material includesluminol; examples of bioluminescent materials include luciferase,luciferin, and aequorin, and examples of suitable radioactive materialinclude ¹²⁵I, ¹³¹I, ³⁵S or ³H.

Antibodies of the invention, including polyclonal, monoclonal, humanizedand fully human antibodies, may be used as therapeutic agents. Suchagents will generally be employed to treat or prevent a disease orpathology associated with aberrant expression or activation of a giventarget in a subject. An antibody preparation, preferably one having highspecificity and high affinity for its target antigen, is administered tothe subject and will generally have an effect due to its binding withthe target. Administration of the antibody may abrogate or inhibit orinterfere with the signaling function of the target. Administration ofthe antibody may abrogate or inhibit or interfere with the binding ofthe target with an endogenous ligand to which it naturally binds. Forexample, the antibody binds to the target and neutralizes or otherwiseinhibits the interaction between CD47 and SIRPα.

A therapeutically effective amount of an antibody of the inventionrelates generally to the amount needed to achieve a therapeuticobjective. As noted above, this may be a binding interaction between theantibody and its target antigen that, in certain cases, interferes withthe functioning of the target. The amount required to be administeredwill furthermore depend on the binding affinity of the antibody for itsspecific antigen, and will also depend on the rate at which anadministered antibody is depleted from the free volume other subject towhich it is administered. Common ranges for therapeutically effectivedosing of an antibody or antibody fragment of the invention may be, byway of nonlimiting example, from about 0.1 mg/kg body weight to about 50mg/kg body weight. Common dosing frequencies may range, for example,from twice daily to once a week.

Antibodies or a fragment thereof of the invention can be administeredfor the treatment of a variety of diseases and disorders in the form ofpharmaceutical compositions. Principles and considerations involved inpreparing such compositions, as well as guidance in the choice ofcomponents are provided, for example, in Remington: The Science AndPractice Of Pharmacy 19th ed. (Alfonso R. Gennaro, et al., editors) MackPub. Co., Easton, Pa.: 1995; Drug Absorption Enhancement: Concepts,Possibilities, Limitations, And Trends, Harwood Academic Publishers,Langhorne, Pa., 1994; and Peptide And Protein Drug Delivery (Advances InParenteral Sciences, Vol. 4), 1991, M. Dekker, New York.

Where antibody fragments are used, the smallest inhibitory fragment thatspecifically binds to the binding domain of the target protein ispreferred. For example, based upon the variable-region sequences of anantibody, peptide molecules can be designed that retain the ability tobind the target protein sequence. Such peptides can be synthesizedchemically and/or produced by recombinant DNA technology. (See, e.g.,Marasco et al., Proc. Natl. Acad. Sci. USA, 90: 7889-7893 (1993)). Theformulation can also contain more than one active compound as necessaryfor the particular indication being treated, preferably those withcomplementary activities that do not adversely affect each other.Alternatively, or in addition, the composition can comprise an agentthat enhances its function, such as, for example, a cytotoxic agent,cytokine, chemotherapeutic agent, or growth-inhibitory agent. Suchmolecules are suitably present in combination in amounts that areeffective for the purpose intended.

The active ingredients can also be entrapped in microcapsules prepared,for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacrylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles, andnanocapsules) or in macroemulsions.

The formulations to be used for in vivo administration must be sterile.This is readily accomplished by filtration through sterile filtrationmembranes.

Sustained-release preparations can be prepared. Suitable examples ofsustained-release preparations include semipermeable matrices of solidhydrophobic polymers containing the antibody, which matrices are in theform of shaped articles, e.g., films, or microcapsules. Examples ofsustained-release matrices include polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradablelactic acid-glycolic acid copolymers such as the LUPRON DEPOT™(injectable microspheres composed of lactic acid-glycolic acid copolymerand leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid. Whilepolymers such as ethylene-vinyl acetate and lactic acid-glycolic acidenable release of molecules for over 100 days, certain hydrogels releaseproteins for shorter time periods.

An antibody according to the invention can be used as an agent fordetecting the presence of a given target (or a protein fragment thereof)in a sample. In some embodiments, the antibody contains a detectablelabel. Antibodies are polyclonal, or more preferably, monoclonal. Anintact antibody, or a fragment thereof (e.g., F_(ab), scFv, orF_((ab)2)) is used. The term “labeled”, with regard to the probe orantibody, is intended to encompass direct labeling of the probe orantibody by coupling (i.e., physically linking) a detectable substanceto the probe or antibody, as well as indirect labeling of the probe orantibody by reactivity with another reagent that is directly labeled.Examples of indirect labeling include detection of a primary antibodyusing a fluorescently-labeled secondary antibody and end-labeling of aDNA probe with biotin such that it can be detected withfluorescently-labeled streptavidin. The term “biological sample” isintended to include tissues, cells and biological fluids isolated from asubject, as well as tissues, cells and fluids present within a subject.Included within the usage of the term “biological sample”, therefore, isblood and a fraction or component of blood including blood serum, bloodplasma, or lymph. That is, the detection method of the invention can beused to detect an analyte mRNA, protein, or genomic DNA in a biologicalsample in vitro as well as in vivo. For example, in vitro techniques fordetection of an analyte mRNA include Northern hybridizations and in situhybridizations. In vitro techniques for detection of an analyte proteininclude enzyme linked immunosorbent assays (ELISAs), Western blots,immunoprecipitations, and immunofluorescence. In vitro techniques fordetection of an analyte genomic DNA include Southern hybridizations.Procedures for conducting immunoassays are described, for example in“ELISA: Theory and Practice: Methods in Molecular Biology”, Vol. 42, J.R. Crowther (Ed.) Human Press, Totowa, N.J., 1995; “Immunoassay”, E.Diamandis and T. Christopoulus, Academic Press, Inc., San Diego, Calif.,1996; and “Practice and Theory of Enzyme Immunoassays”, P. Tijssen,Elsevier Science Publishers, Amsterdam, 1985. Furthermore, in vivotechniques for detection of an analyte protein include introducing intoa subject a labeled anti-analyte protein antibody. For example, theantibody can be labeled with a radioactive marker whose presence andlocation in a subject can be detected by standard imaging techniques.

Pharmaceutical Compositions

The antibodies of the invention (also referred to herein as “activecompounds”), and derivatives, fragments, analogs and homologs thereof,can be incorporated into pharmaceutical compositions suitable foradministration. Such compositions typically comprise the antibody and apharmaceutically acceptable carrier. As used herein, the term“pharmaceutically acceptable carrier” is intended to include any and allsolvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like,compatible with pharmaceutical administration. Suitable carriers aredescribed in the most recent edition of Remington's PharmaceuticalSciences, a standard reference text in the field, which is incorporatedherein by reference. Preferred examples of such carriers or diluentsinclude, but are not limited to, water, saline, ringer's solutions,dextrose solution, and 5% human serum albumin. Liposomes and non-aqueousvehicles such as fixed oils may also be used. The use of such media andagents for pharmaceutically active substances is well known in the art.Except insofar as any conventional media or agent is incompatible withthe active compound, use thereof in the compositions is contemplated.Supplementary active compounds can also be incorporated into thecompositions.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid(EDTA); buffers such as acetates, citrates or phosphates, and agents forthe adjustment of tonicity such as sodium chloride or dextrose. The pHcan be adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CREMOPHOREL™ (non-ionic oil-in-water emulsifier and/or solubilizer) (BASF,Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, thecomposition must be sterile and should be fluid to the extent that easysyringeability exists. It must be stable under the conditions ofmanufacture and storage and must be preserved against the contaminatingaction of microorganisms such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (for example, glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), and suitable mixtures thereof. The properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, polyalcohols such asmanitol, sorbitol, sodium chloride in the composition. Prolongedabsorption of the injectable compositions can be brought about byincluding in the composition an agent which delays absorption, forexample, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound in the required amount in an appropriate solvent with one or acombination of ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the active compound into a sterile vehicle that contains abasic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, methods of preparation are vacuum dryingand freeze-drying that yields a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed. Pharmaceutically compatible binding agents, and/or adjuvantmaterials can be included as part of the composition. The tablets,pills, capsules, troches and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orSterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g.,with conventional suppository bases such as cocoa butter and otherglycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

The invention will be further described in the following examples, whichdo not limit the scope of the invention described in the claims.

EXAMPLES Example 1: Cloning, Expression and Purification of Human CD47

Cloning.

The sequence corresponding to the extracellular domain of human CD47(hCD47), was amplified from human CDNA by polymerase chain reaction(PCR) using specific oligonucleotides. The amplification production wasgel-purified and cloned into the Peak8 mammalian expression vector (EdgeBiosystems, Gaithersburg, Md.). The vector was further modified tointroduce an AVITAG™ (biotin-conjugated peptide tag) (Avidity, DenverColo.) and an hexa-histidine tag at the C-terminus allowing for singlesite biotinylation of the protein and purification by IMAC (ImmobilizedMetal Ion Affinity Chromatography). The constructs were verified by DNAsequencing.

Expression.

The plasmid was then transfected into mammalian cells using a liposomebased transfection reagent such as TransIT-LT1 (Mirus, Madison, Wis.).The transfection step requires only small quantities of DNA and cells,typically 2×10⁵ cells and 2 μg of plasmid DNA per well and thetransfection carried out in a 6-well plate. Although different mammaliancell lines can be used, in the examples given below, transformed humanembryo kidney monolayer epithelial cells (PEAK cells) were transfected.These cells stably express the EBNA-1 gene, further supporting theepisomal replication process, are semi-adherent and can be grown understandard conditions cell culture incubator (5% CO2; 37° C. in DMEMmedium supplemented with 10% fetal calf serum). After 24 h, cells wereplaced under selective conditions by adding medium containing 0.5-2μg/mL puromycin, as cells harboring the episomal vector are resistant tothis antibiotic.

Two to three weeks after transfection, were used to seed Tri-flasks ordisposable CELLLINE™ bioreactors (membrane cell culture flask) for theproduction step. The CELLLINE™ is a two compartment bioreactor that canbe used in a standard cell culture incubator. The smaller compartment(15 ml) contains the cells and is separated from a larger (one liter)medium containing compartment by a semi-permeable membrane with acut-off size of 10 kDa (Bruce et al. 2002, McDonald et al. 2005). Thissystem allows for the diffusion of nutrients, gazes and metabolic wasteproducts, while retaining cells and secreted proteins in the smallercompartment. The culture was maintained for 7-10 days before harvest ofthe supernatant. As the medium contains serum, the cells maintain goodviability and several production runs can be generated using the samecells and containers.

Purification.

After harvest, the cell culture supernatants were clarified bycentrifugation and filtered through a 0.22 μm membrane. The supernatantfrom Tri-flasks were concentrated 20-40 times using a concentrationdevice such as a SartoFlow 200 (Sartorius) with a membrane having anappropriate cut-off size to retain the protein of interest. This stepwas not required using the CELLline bioreactor due to the low volumerecovered from the cell compartment. In addition, the concentration stepincreases the concentration of both the protein of interest and highmolecular weight contaminants such as bovine serum albumin orimmunoglobulins. In contrast, the supernatant retrieved from the cellcompartment of the CELLine bioreactor contains concentrated recombinantprotein and reduced levels of contaminants as they cannot cross the 10kDa membrane separating the two chambers of the reactor. This increasedrecombinant protein to contaminant ratio greatly enhances the efficiencyof purification using IMAC. The concentrated supernatant was thensupplemented with 100 mM imidazole and loaded on Ni-NTA affinitychromatography resin (Qiagen). The relatively high concentration ofimidazole minimizes binding of contaminants to the resin. After washingof the column, proteins are eluted at a flow rate of 2 mL/min using a 30mL imidazole gradient (20-400 mM imidazole) on an AKTA Primechromatography system (Amersham Pharmacia Biotech). The elution gradientfurther improves the purity of the recombinant protein but can bereplaced by a step elution approach if a chromatography system is notavailable. The eluted fractions can be analyzed by SDS-PAGE or ELISA todetermine their content in recombinant protein. The fractions ofinterest are pooled and desalted on PD-10 columns (GE Healthcare)equilibrated with phosphate buffered saline or another appropriatebuffer. The desalted proteins can then be quantified using varioustechniques and their purity analyzed by SDS-PAGE. Recombinant CD47 wasbiotinylated in vitro using biotin ligase (Avidity, Denver Colo.)according to manufacturer's instructions. After desalting thebiotinylation level was evaluated by pull-down assays using streptavidinmagnetic beads and SDS-PAGE analysis.

Example 2: Cloning, Expression and Purification of Human CD19

Cloning.

The sequence corresponding to the extracellular domain of human CD19(hCD419), was amplified from human cDNA by polymerase chain reaction(PCR) using specific oligonucleotides. The amplification production wasgel-purified and cloned into the pEAK8 mammalian expression vector (EdgeBiosystems, Gaithersburg, Md.). The vector was further modified tointroduce an Avitag™ (Avidity, Denver Colo.) and an hexa-histidine tagat the C-terminus allowing for single site biotinylation of the proteinand purification by IMAC (Immobilized Metal Ion AffinityChromatography). The constructs were verified by DNA sequencing.

Expression and Purification.

The expression, purification and biotinylation of soluble hCD19 wereperformed as described in Example 1.

Example 3: Phage Display Selection Using Human scFv Libraries ContainingFixed Variable Heavy Chain

General procedures for construction and handling of human scFv librariesdisplayed on M13 bacteriophage are described in Vaughan et al., (Nat.Biotech. 1996, 14:309-314), hereby incorporated by reference in itsentirety. The libraries used for selection and screening encode scFvthat all share the same VH domain and are solely diversified in the VLdomain. Methods for the generation of fixed VH libraries and their usefor the identification and assembly of bispecific antibodies aredescribed in US 2012/0184716 and WO 2012/023053, each of which is herebyincorporated by reference in its entirety. The procedures to identifyscFv binding to hCD19 or hCD47 are described below.

Liquid Phase Selections.

Aliquots of scFv phage libraries (10¹² Pfu) were blocked with PBScontaining 3% (w/v) skimmed milk for one hour at room temperature on arotary mixer. Blocked phage was then deselected on streptavidin magneticbeads (Dynal M-280) for one hour at room temperature on a rotary mixer.Deselected phage was then incubated with in vivo biotinylated hCD19 orhCD4 7 (100 nM) for two hours at room temperature on a rotary mixer.Beads were captured using a magnetic stand followed by four washes withPBS/0.1% TWEEN® 20 (polysorbate 20) and 3 washes with PBS. Beads werethen directly added to 10 ml of exponentially growing TG1 cells andincubated for one hour at 37° C. with slow shaking (100 rpm). An aliquotof the infected TG1 was serial diluted to titer the selection output.The remaining infected TG1 were spun at 3000 rpm for 15 minutes andresuspended in 0.5 ml 2×TY-AG (2×TY media containing 100 μg/mlampicillin and 2% glucose) and spread on 2×TY AG agar Bioassay plates.After overnight incubation at 30° C. 10 ml of 2×TY AG was added to theplates and the cells were scraped form the surface and transferred to a50 ml polypropylene tube. 2×TYAG containing 50% glycerol was added tothe cell suspension to obtain a final concentration of 17% glycerol.Aliquots of the selection round were kept at −80° C.

Phage Rescue.

100 μl of cell suspension obtained from previous selection rounds wereadded to 20 ml of 2×TYAG and grown at 37° C. with agitation (240 rpm)until an OD600 of 0.3 to 0.5 was reached. The culture was thensuper-infected with 3.3×10¹⁰ MK13K07 helper phage and incubated for onehour at 37° C. (150 rpm). The medium was then changed by centrifugingthe cells at 2000 rpm for 10 minutes, removing the medium andresuspending the pellet in 20 ml of 2×TY-AK (100 μg/ml ampicillin; 50μg/ml kanamycin). The culture was then grown overnight at 30° C. (240rpm). The next day, the phage containing supernatant was used for thenext round of selection.

Cell Surface Selections.

Phage containing supernatants were blocked with PBS containing 3% (w/v)skimmed milk for one hour at room temperature on a rotary mixer. Blockedphage was then deselected for one hour on Jukat T cells that do notexpress CD19 and that had been previously blocked with PBS containing 2%(w/v) skimmed milk. Deselected phage was then incubated with 2×10⁷ Rajicells expressing CD19 for one hour at room temperature with gentleshaking. Cells were then pelleted and washed ten times with PBS. Boundphage was eluted by adding directly 10 ml of exponentially growing TG1to the T75 flask and incubating for one hour at 37° C. with slowshaking. An aliquot of the infected TG1 was serial diluted to titer theselection output. Infected TG1 were spun at 3000 rpm for 15 minutes andre-suspended in 0.5 ml 2×TY-AG (2×TY media containing 100 μg/mlampicillin and 2% glucose) and spread on 2×TYAG agar Bioassay plates.After overnight incubation at 30° C. 10 ml of 2×TYAG was added to theplates and the cells were scraped form the surface and transferred to a50 ml polypropylene tube. 2×TYAG containing 50% glycerol was added tothe cell suspension to obtain a final concentration of 17% glycerol.Aliquots of the selection round were kept at −80° C.

scFv Periplasmic Preparation for Binding and Functional Tests.

Individual clones were inoculated into a deep well microtiter platecontaining 0.9 ml of 2×TYAG media (0.1% glucose) per well and grown at37° C. for 5-6 h (250 rpm). 100 μl per well of 0.2 mM IPTG in 2×TYmedium were then added to give a final concentration of 0.02 mM IPTG.Plates were then incubated overnight at 30° C. with shaking at 250 rpm.The deepwell plates were centrifuged at 2,500 rpm for 10 min and thesupernatant carefully removed. The pellets were re-suspended in 150 μlTES buffer (50 mM Tris/HCl (pH 8), 1 mM EDTA (pH 8), 20% sucrose,complemented with COMPLETE™ protease inhibitor (protease inhibitorcocktail tablet), Roche). A hypotonic shock was produced by adding 150μl of diluted TES buffer (1:5 TES:water dilution) and incubation on icefor 30 min. Plates were then centrifuged at 4000 rpm for 10 minutes toremove cells and debris. The supernatants were carefully transferredinto another microtiter plate and kept on ice for immediate testing infunctional assays or binding assays.

Phage Clone Sequencing.

Single clones were placed in a microtiter plate containing 150 μl of2×TYAG media (2% glucose) per well and grown at 30° C. (120 rpm)overnight. The next day 5 μl of culture was transferred into anotherplate containing 45 μl of dH2O and mixed. The plate was then frozen at−20° C. After thawing, 1 μl of this suspension was used for PCRamplification using standard PCR protocols with primer specific forpNDS1: mycseq, 5′-CTCTTCTGAGATGAGTTTTTG-3′ (SEQ ID NO: 283) andgene3leader, 5′-TTATTATTCGCAATTCCTTTAGTTGTTCCT-3′ (SEQ ID NO: 284). ThePCR reactions were purified in 96 well format using the Montage PCR 96system (Millipore). 5 μl of the eluted DNA was sequencing using themycseq and gene3leader primers.

Example 4: Screening for scFv Binding to hCD47 and scFv Inhibiting SIRPαInteraction

Binding:

Screening of scFv for binding to hCD47 was tested in a homogenous assayusing FMAT technology. The following reagents were mixed in each well ofa 384 optical plate (Costar): 30 μl of a streptavidin polystyrene beadsuspension (Spherotech; 3000 beads/well) coated with biotinylated hCD47or a control biotinylated protein (NusA); 60 μl of scFv periplasmicpreparation; 10 μl of detection buffer (PBS containing anti-cmycantibody at 5 μg/mL; anti-mouse FC ALEXAFLUOR® 647 (photostablefluorescent dye) diluted 1:200). After mixing at 450 rpm for 5 minutes,the 384 well plates were incubated at room temperature and read after 1and 3 hours on an FMAT 8200 Cellular Detection System (AppliedBiosystems). Each scFv sample was tested in duplicate against hCD47 andNusA. Clones expressing scFv giving a specific signal for hCD47 and notNusA were selected for further analysis.

Inhibition of CD47-SIRPα Interaction:

ScFv were also screened for their capacity to inhibit the interactionbetween CD47 and SIRPα in a bead based homogenous assay using FMATtechnology. Protein A polystyrene beads (Spherotech) are incubated with5 μg/mL of goat anti-human lgG Fcγ specific (Jackson Immunoresearch).After washing of the beads 5 μg/mL SIRPα-Fc (R&D Systems) was added sothat the fusion protein can be captured at the bead surface. Afterblocking with PBS; 2% Tropix 1-block (Applied Biosystems), 30 μl of thebeads suspension (3000 beads/well) coated were added to each well of a384 optical plate (Costar). In a separate 96 well plate 120 μl of scFvperiplasmic preparation were mixed with 24 μl of biotinylated hCD47 (300ng/ml) and incubated for 50 minutes at room temperature so that the scFvcan bind to hCD47. After incubation, 24 μl of Streptavidin Cy5 (1 μg/ml;Invitrogen) are added to the mix and 70 μl of this final mix are addedto the 30 μl of beads in each well of the 384 well plate. After 3 hoursof incubation at room temperature, the plate is then read on an FMAT8200 Cellular Detection System (Applied Biosystems). Controls wellcontaining no scFv or an irrelevant scFv not binding to CD47 wereincluded in each plate so that clones expressing scFv leading to areduction of the SIRPα-CD47 signal measured in controls were selectedfor further analysis.

Alternatively a cell based assay monitoring the interaction of solubleSIRPα with hCD47 expressed at the surface of stably transfected Chinesehamster ovary (CHO) cell line was also used for screening of candidates.20 μl of PBS-BSA 2% azide 0.1% containing 3000 CHO expressing hCD47cells were added to each well of a 384 optical plate (Costar). 50 μl ofa twofold dilution of the scFv periplasmic preparation was then added tothe well and incubated at room temperature for 30 minutes to allow thescFv to bind to CD47 on cells. After incubation 30 μl of PBS 2% BSAazide 0.1% containing 10 ng/ml of SIRPα-Fc (R&D systems) and AntihIgG-Fc FMAT Blue coupled antibody (diluted 1:2000) were added andfurther incubated for 3 hours before reading on an FMAT 8200 CellularDetection System (Applied Biosystems).

Example 5: Screening for scFv Binding to hCD19

Screening of scFv for binding to recombinant hCD19 was tested in ahomogenous assay using FMAT technology as described in Example 4.

Screening was also performed on Raji cells for binding to the nativeform of hCD19. To each well of a 384 optical plate (Costar) 30 μl ofPBS-BSA 2% azide 0.1% containing 3000 Raji cells (a human B cell lineexpressing CD19) or Jurkat cells (a human T cell line that do notexpress CD19) were added. Then, 30 μl of a twofold dilution of the scFvperiplasmic preparation, 30 μl of PBS-BSA2% and 10 30 μl of 10×detection buffer (Qiagen Antibody pentaHis AF647 diluted 1:700 inPBS-BSA2%). After mixing at 450 rpm for 5 minutes, the 384 well plateswere incubated at room temperature and read after 1 and 3 hours on anFMAT 8200 Cellular Detection System (Applied Biosystems). Clonesexpressing scFv giving a specific signal for Raji and not Jurkat cellswere selected for further analysis.

Example 6: Fixed VH Candidates Reformatting into IgG and TransientExpression in Mammalian Cells

After screening, scFv candidates against hCD19 or hCD47 were reformattedinto IgG and expressed by transient transfection into PEAK cells. The VHand VL sequences of selected scFv were amplified with specificoligonucleotides and cloned into an expression vector containing theheavy and light chain constant regions and the constructions wereverified by sequencing. The expression vectors were transfected intomammalian cells using the Fugene 6 Transfection Reagent (Roche, Basel,Switzerland). Briefly, Peak cells were cultured in 6-well plates at aconcentration of 6×10⁵ cells per well in 2 ml culture media containingfetal bovine serum. The expression vectors encoding the candidate VH andVL sequences were co-transfected into the cells using the Fugene 6Transfection Reagent according to manufacturer's instructions. One dayfollowing transfection, the culture media was aspirated, and 3 ml offresh serum-free media was added to cells and cultured for three days at37° C. Following three days culture period, the supernatant washarvested for IgG purified on protein G-Sepharose 4B fast flow columns(Sigma, St. Louis, Mo.) according to manufacturer's instructions.Briefly, supernatants from transfected cells were incubated overnight at4° C. with ImmunoPure (G) IgG binding buffer (Pierce, Rockford Ill.).Samples were then passed over Protein G-Sepharose 4B fast flow columnsand the IgG consequently purified using elution buffer. The eluted IgGfraction was then dialyzed against PB S and the IgG content quantifiedby absorption at 280 nm. Purity and IgG integrity were verified bySDS-PAGE.

Example 7: Affinity Modulation of Anti-hCD47 Antibodies (a) AntibodiesKa3, Ke8, Kc4

Three antibodies identified during the screening process described inthe Examples above were shown to be specific for human CD47 and able toblock the interaction between CD47 and SIRPα were selected for affinitymaturation in order to increase their affinity and potency. All theseantibodies share the same variable heavy chain but have differentvariable light chains. Ka3 and Ke8 contain a kappa light chain (IGVK1-39according to the IMGT nomenclature) whereas Kc4 contains a lambda lightchain (IGVL2-14). Several phage libraries displaying scFv variants weregenerated by introducing diversity into the CDR1, CDR2 and CDR3 of thevariable light chain region while the heavy chain variable region waskept unmodified. One library of 9×10⁷ transformants covering atheoretical diversity of 7×10⁵ was generated for Ka3; two libraries of2×10⁸ transformants each, partially covering a theoretical diversity of2.4×10⁹ were generated for Ke8 and one library of 3.6×10⁷ transformantscovering a theoretical diversity of 2.6×10⁵ was generated for Kc4. Theselibraries were used for phage display selections as described in Example3 except that the selection stringency was increased between rounds byreducing the concentration of hCD47 between different rounds: 10 nM and1 nM of hCD47 were used in the first and second round of selection,respectively. The selected variants were screened for the capacity toinhibit the interaction between hCD47 and SIRPα using the assaydescribed in Example 4. Positive clones were then reformatted as IgG andcharacterized as described in the following Examples. These affinitymaturation efforts lead to the identification of the following anti-VD47antibodies:

-   -   Ke8H6; Ke86G9; Ke8A3; Ke8C4; Ke8F1; Ke8B7; Ke8G11; Ke8A8; Ke8A4;        Ke8B2; Ke8C7; Ke8H3; Ke8A2; Ke8H5; Ke8G6; Ke8E8; Ke81A3; Ke81G9;        Ke84G9; Ke8G2; Ke8F2    -   Ka3G2; Ka3D3; Ka3A2; Ka3B2; Ka3C5; Ka3A3; Ka3H8; Ka3H3    -   Kc4E2; Kc4F4; Kc4A1; Kc4C11; Kc4E10; Kc4B1; Kc4C3; Kc4A4;        Kc4G11; Kc4G9

(b) 5A3 Antibody Engineering for Affinity Modulation

The VL sequence of anti-CD47 5A3 antibody was engineered to decrease itsaffinity toward its target. The 5A3-VL sequence was aligned to itsclosest germline sequence, the human IGKV1-33 according to the IMGTnomenclature (FIG. 1). Using this alignment, several residues wereidentified in the CDRL1 and CDRL2 of 5A3 VL which are not conserved withthe germline sequence. Some of these amino acids were mutated in orderto alter the binding affinity of the antibody. Residues of the 5A3 CDRL3were also changed to modulate antibody binding while at the same timetargeting the same epitope on CD47. These different strategies led tothe identification of the 5A3-M3 and 5A3-M5 candidates.

These variants were first tested in a CD47/SIRPα binding assay todetermine their blocking potency compared to the parental 5A3 antibody(FIG. 2). The 5A3-M3 and 5A3-M5 are both less potent at inhibiting theinteraction between CD47 and SIRPα than 5A3 with 5A3-M5 showing theweakest inhibition potency profile.

The affinity of these variants for human CD47 was then evaluated bysurface plasmon resonance technology. The K_(D) of the 5A3, 5A3-M3 and5A3-M5 antibodies are about 2.36E-08, 5.60E-08 and 2.84E-06 M,respectively. These data confirmed that the 5A3 variants are binding toCD47 with a lower affinity compared to the parental antibody and thatthe 5A3-M5 has the weakest affinity for human CD47 while the 5A3-M3 hasan intermediate affinity.

Example 8: Characterization of CD47 Antibodies

Binding of CD47 Antibodies to huCD47-Transfected CHO Cells

The specificity of CD47 monoclonal antibodies (Mabs) was shown by flowcytometry using CHO cells stably transfected with human CD47 (CHO-huCD47cells). Non-transfected CHO cells were used as control. In brief,purified CD47 Mabs were incubated with CHO-huCD47 cells at a finalconcentration of 10 μg/ml for 30 minutes. After two washes, bound CD47antibodies were detected using a Cy-5 conjugated anti-human Fc secondaryantibody (BD biosciences). FIG. 3 shows a significant binding of CD47MAbs to hu-CD47 transfected CHO cells, but no binding (orbackground-level binding) to non-transfected CHO cells, thusdemonstrating the specificity of CD47 MAbs of the present invention.

Binding of CD47 Antibodies to HEK293-P Cells

The specificity of CD47 Mabs was further confirmed in an experimentusing HEK293-P cells with a siRNA mediated CD47 gene knock-down. TheHEK293-P cells (Peak cells) were derived from human embryonic kidneycells and expresses low to moderate levels of CD47. A CD47-deficientvariant of Peak cells has been generated by stably transfecting themwith siRNA specific to the CD47 gene. Cell surface expression of CD47antigen is reduced in these CD47 knock-down PEAK cells by more than 85%(data not shown). FIG. 4 demonstrates the binding of selected CD47 MAbsto non-transfected Peak cells and to CD47 knock-down Peak cells. Bindingof CD47 Mabs to CD47 siRNA-transfected Peak cells is significantlyreduced, thus confirming their antigen specificity.

Cross-Reactivity of CD47 Antibodies with Cynomolgus CD47; Binding ofCD47 Antibodies to Human and Cynomolgus CD4+ T Cells

The ability of CD47 monoclonal antibodies of the present invention tocross-react with native cynomolgus monkey CD47 was tested by flowcytometry. Binding of CD47 antibodies to cynomolgus CD4-positive Tlymphocytes present in peripheral blood mononuclear cells (PBMCs) wascompared to the binding to the corresponding human cell population. Inbrief, cynomolgus peripheral blood mononuclear cells (PBMCs) wereobtained from Ricerca Biosciences. Human PBMCs were isolated from abuffy coat using CPT ficoll tubes (Beckton and Dickinson). For flowcytometry analysis, PBMCs were preincubated with FcgR Blocking Reagent,(Miltenyi Biotech.) for 20 minutes in order to block Fe gamma receptorsbefore addition of CD47 antibodies (final concentration of 0.005 mg/ml).After an incubation period of 30 minutes cells were washed and reactedwith PE-conjugated anti human CD4 antibody (clone L200, BD Pharmingendiluted 1/100) and FMAT-BLUE®-conjugated (monofunctional dye label)goat-anti human Fe antibody (Jackson Immuno Research, 109-005-098). TheMFI for CD47 binding (FL4) was then determined by flow cytometry in theCD4+ positive population (gated on FL2). As shown in FIG. 5, CD47monoclonal antibodies of the present invention bind to native human CD47and cross-react with cynomolgus CD47.

SIRPα Blocking Activity of CD47 Antibodies

The SIRPα blocking activity of CD47 was determined in the CD47-SIRPαcompetitive binding assay. Dose-response experiments with CD47 Mabsallowed determining an IC50 value for each of the CD47 MAbs of thepresent invention. In brief, human CD47 transfected CHO cells wereincubated with His-tagged soluble human SIRPα (final concentration, 200ng/ml) and increasing concentrations of CD47 Mab (3.3 pM to 330 nM, inquadruplicates) The detection of bound SIRPα was as described in example4. FIG. 6 shows the potency of CD47 Mabs to block the CD47-SIRPαinteraction, represented by IC50 values. CD47 Mabs are grouped by familyand ranked from higher to lower potency. Their neutralizing activity wascompared to the commercially available CD47 antibody B6H12. It isapparent from FIG. 6 that the neutralizing potencies of CD47 Mabs of thepresent invention vary over a wide range.

Hemagglutination Activity of CD47 Antibodies

FIG. 7 demonstrates that high-affinity CD47 Mabs of the 5A3, Ke8, andKa3 families induce hemagglutination; in contrast to the other threefamilies, Kc4 family antibodies tested in this experiment do not seem toinduce hemagglutination even the one binding strongly to CD47 andinhibiting potently the CD47-SIRPα interaction.

CD47 MAbs were tested for their ability to induce homotypic clusteringof erythrocytes (hemagglutination). 10 microliters of human whole bloodwas diluted in 40 microliters of antibody solution in PBS at differentconcentrations (range: 0.003 microg/ml to 50 microg/ml final Mabconcentration) in flat-bottom 96 well plates. The blood-antibody mix wasincubated O/N at 37° C. without shaking. At the end of the incubation,the plates were agitated manually, tilted at about 30° C., and let torest for about 10 minutes.

Evidence of hemagglutination is demonstrated by the formation of aclumped deposit, in the form of a crescent at the bottom around theinferior border of the well. All but the lowest affinity CD47 antibodiesof the 5A3, Ke8, and Ka3 families (specifically, 5A3M5, Ke8A3, Ka3A3)caused hemagglutination. In contrast, the CD47 antibodies of the Kc4family did not cause hemagglutination, even the higher affinity ones(Kc4E2, Kc4F4).

Example 9: CD19 Antibody Affinity Maturation (a) Antibody B7

Amongst the antibodies identified during the screening process describedin the Examples above B7 was selected for affinity maturation in orderto increase its affinity for hCD19. Candidate B7 contains a lambda lightchain (IGLV6-57) and several phage libraries displaying scFv variantswere generated by introducing diversity into the CDR1, CDR2 and CDR3 ofthe variable light chain region while the heavy chain variable regionwas kept unmodified. Different diversification strategies were used togenerate 20 libraries comprising a total of 2×10⁹ transformantspartially covering a theoretical diversity of 4×10¹².

(b) Antibody L7B7_D11

Antibody D11 was identified during the affinity maturation of B7described above and binds to hCD19 with a higher affinity than theparental antibody B7. This antibody was selected for a second round ofaffinity maturation of its light chain. A total of 6 librariescomprising 2.8×10⁹ transformants partially covering a theoreticaldiversity of 4×10⁹ were generated and used for phage display selectionsas described above except that 1 nM of hCD19 was used for each round ofselection. This second round of affinity maturation lead to theidentification of the following antibodies with and improved binding toCD19: L7B7_C2; L7B7_A6; L7B7_Bll; L7B7 C6 and L7B7_C9.

Example 10: Expression and Purification of Bispecific AntibodiesCarrying a Lambda and a Kappa Light Chain

The simultaneous expression of one heavy chain and two lights chain inthe same cell can lead to the assembly of three different antibodies.Simultaneous expression can be achieved in different ways such as thatthe transfection of multiple vectors expressing one of the chains to beco-expressed or by using vectors that drive multiple gene expression. Avector pNovi κHλ was previously generated to allow for the co-expressionof one heavy chain, one Kappa light chain and one Lambda light chain asdescribed in US 2012/0184716 and WO 2012/023053, each of which is herebyincorporated by reference in its entirety. The expression of the threegenes is driven by human cytomegalovirus promoters (hCMV) and the vectoralso contains a glutamine synthetase gene (GS) that enables theselection and establishment of stable cell lines. The VH and VL gene ofthe anti-hCD19 IgGλ or the anti-hCD47 IgGκ were cloned in the vectorpNovi κHλ, for transient expression in mammalian cells. Peak cells werecultured in 6-well plates at a concentration of 6×10⁵ cells per well in2 ml culture media containing fetal bovine serum. 2 μg of plasmid DNAwas transfected into the cells using TransIT-LT1 transfection reagent(Mirus) according to manufacturer's instructions. Antibody concentrationin the serum-containing supernatant of transfected cells was measured atseveral time points during the production using the Bio-LayerInterferometry (BLI) technology. An OctetRED96 instrument and Protein Abiosensors were used for quantitation (Pall, Basel, Switzerland). 200 μLof supernatant were used to determine IgG concentration; biosensors werepre-conditioned and regenerated using 10 mM glycine pH 1.7 and IgGcalibrators diluted in conditioned PEAK cell medium were prepared forstandard curve generation. Concentrations were determined using the doseresponse 5PL weighted Y standard curve equation and an initial slopebinding rate equation. According to antibody concentration, supernatantswere harvested 7 to 10 days after transfection and clarified bycentrifugation at 1300 g for 10 min. The purification process wascomposed of three affinity steps. First, the CaptureSelect™ IgG-CH1affinity matrix (Life Technologies, Zug, Switzerland) was washed withPBS and then added in the clarified supernatant. After incubationovernight at +4° C., supernatants were centrifuged at 1000 g for 10 min,flow through was stored and resin washed twice with PBS. Then, the resinwas transferred on spin columns and a solution containing 50 mM glycineat pH 2.7 was used for elution. Several elution fractions weregenerated, pooled and desalted against PBS using 50 kDa Amicon® UltraCentrifugal filter units (Merck KGaA, Darmstadt, Germany). The finalproduct, containing total human IgGs from the supernatant, wasquantified using a Nanodrop spectrophotometer (NanoDrop Technologies,Wilmington, Del.) and incubated for 15 min at RT and 20 rpm with theappropriate volume of CaptureSelect™ LC-kappa (Hu) affinity matrix (LifeTechnologies, Zug, Switzerland). Incubation, resin recovery, elution anddesalting steps were performed as described previously. The lastaffinity purification step was performed using the CaptureSelect™LC-lambda (Hu) affinity matrix (Life Technologies, Zug, Switzerland)applying the same process as for the two previous purifications. Thefinal product was quantified using the Nanodrop. Purified bispecificantibodies were analyzed by electrophoresis in denaturing and reducingconditions. The Agilent 2100 Bioanalyzer was used with the Protein 80kit as described by the manufacturer (Agilent Technologies, Santa Clara,Calif., USA). 4 μL of purified samples were mixed with sample buffersupplemented with dithiothreitol (DTT; Sigma Aldrich, St. Louis, Mo.).Samples were heated at 95° C. for 5 min and then loaded on the chip. Allsamples were tested for endotoxin contamination using the LimulusAmebocyte Lysate test (LAL; Charles River Laboratories, Wilmington,Mass.).

Example 11: Characterization of Monovalent and Bispecific Antibodies

Dual-targeting bispecific antibodies bind to two different antigens onthe surface of the same cell. Simultaneous binding of the two antibodyarms to two antigens on the surface of the cell (termed co-engagement)results in additive or synergistic increase of affinity due to aviditymechanism. As a consequence, co-engagement confers high selectivitytowards cells expressing both antigens as compared to cells that expressjust one single antigen. In addition, the affinities of the two arms ofa bispecific antibody to their respective targets can be set up in a waythat binding to target cells is principally driven by one of theantibody arms. For instance, a dual targeting κλ-body composed of onearm binding with high affinity to a tumor associated antigen (TAA), forexample CD19, and a second arm binding with lower affinity to CD47—butsufficient to inhibit CD47/SIRPα upon TAA co-engagement—should allowpreferential inhibition of CD47 in cancer versus normal cells. Theexperiments described below (FIGS. 9 to 13) compare the bindingaffinity, the CD47-SIRPα neutralization potency, and the tumor cellkilling activity of CD47xCD19 bispecific K-body and the correspondingmonovalent antibody, i.e., having the same CD47-binding arm plus a“dummy” non-binding arm.

Binding of Monovalent and Bispecific Antibodies to B Cell Lines

To demonstrate that binding of CD47xCD19 κλ bodies to target cells isCD19 dependent, a series of FACS experiments comparing the binding ofCD47xCD19 κλ bodies to their monovalent counterparts were performed. Twotypes of cells were used, a CD19-positive Burkitt lymphoma cell lineRaji (expressing about 65,000 CD47 molecules per cell) and theCD19-negative B-NHL cell line DS-1 (expressing about 150,000 CD47molecules per cell) as a control. FIGS. 9A-9C demonstrate that aCD47xCD19 κλ body co-engages the two targets at the surface of Rajicells. This is shown by (i) increased affinity to Raji cells as comparedto DS-1 cells and (ii) increased affinity of the CD47xCD19 κλ body ascompared to the CD47 monovalent antibody, observed with Raji cells—butnot with DS-1 cells. A comparison of FACS profiles generated with thebinding of CD19 monovalent antibody, the CD47 monovalent antibody, andthe CD47xCD19 κλ body to Raji cells clearly demonstrates that binding ofthe CD47xCD19 κλ to target cells is principally driven by the CD19 arm.

SIRPα Blocking Activity of Monovalent and Bispecific Antibodies

Another series of experiments provides a further proof of co-engagementof CD19 and CD47 on the surface of the target cell by showing that theneutralization of CD47-SIRPα interaction by CD47xCD19 κλ bodies isCD19-dependent. In this experiment, the activity of CD47xCD19 κλ bodiesand the corresponding monovalent antibodies was tested in the CD47-SIRPαinhibition assay as described in Example 4. FIG. 10 shows that CD47xCD19κλ bodies inhibited the CD47-SIRPα interaction in Raji cells with asignificantly higher potency than the corresponding CD47 monovalentantibodies. Efficient neutralization of CD47-SIRPα interaction requiredCD19 co-engagement. IC50 values obtained with CD47xCD19 BsAbs are 20 to1000× lower than the values obtained with the corresponding CD47monovalent antibody (see Table 4).

TABLE 4 IC50 values of CD47 monovalent and bispecific antibodies IC50CD47-SIRPα Assay (Raji) CD47 CD47Xcd19 BsAb CD47 MonovalentMonovalent/BsAb Arm [nM] [nM] ratio 5A3M3 0.031 13 419 5A3M4 0.36 4001111 Ke8G11 0.066 1.2 18 Ke8C4 0.12 13 108 Ke8A3 1.1 >500 >500 Ka3G20.11 5.1 46 Ka3 0.32 6.7 21 Ka3H3 0.71 44 62

Example 12: ADCC Mediated by Bispecific Antibodies is CD19-Dependent

The ability of dual targeting κλ-bodies to co-engage CD47 and CD19results in a significant increase in the affinity of binding toCD19-positive cells and in CD19-dependent neutralization of theCD47-SIRPα interaction. This, in turn, translates into efficient andselective cancer cell killing mediated by CD47xCD19 κλ body, asdemonstrated in ADCC and ADCP experiments described in this and thefollowing example.

ADCC assays were performed with unfractionated human PBMC and Raji orRamos B cell lymphoma target cells. Dose-response experiments shown inFIG. 11 demonstrate that CD47xCD19 κλ bodies provided herein kill B celllymphoma cells in a more efficient way than the corresponding CD47monovalent antibodies. Efficient ADCC is therefore dependent on CD19co-engagement. FIG. 11C shows that the efficacy of ADCC with CD47xCD19κλ bodies is comparable to rituximab and that it is significantly higherthan with the CD19 Mab C2.

Example 13: ADCP Mediated by Bispecific Antibodies is CD19-Dependent

FIG. 12 demonstrates that CD47xCD19 BsAbs provided herein phagocytoseCD19-positive cells in a CD19-dependent manner, as the correspondingCD47 monovalent antibodies are much less efficient (if any).

ADCP experiments were performed with human macrophages differentiatedfrom peripheral blood monocytes and Raji as target cells. Macrophageswere co-incubated with CFSE-labeled Raji cells (effector: target ratio1:5) for 2.5 hours at 37° C. in the presence of increasingconcentrations of bispecific or monovalent antibody. At the end of theincubation period, biotinylated anti-human CD14 antibody and Strep-Cy5were added to label the macrophages. The cells were then washed andsubjected to FACS analysis. Phagocytosis was evidenced bydouble-positive events.

Dose-response experiments shown in FIG. 12 demonstrate that CD47xCD19 κλbodies are more potent than the corresponding CD47 monovalentantibodies. Efficient ADCC is therefore dependent on CD19 co-engagement.CD19 co-engagement by the bispecific antibody drives efficacy. What ismore, the experiments shown in FIG. 12 confirm that blocking CD47 isnecessary to elicit efficient ADCP, as the CD19 Mab C2, which bindstarget cells with high affinity, does not induce significantphagocytosis.

Example 14: In Vivo Antitumor Activity of Bispecific Antibodies

The anti-tumor activity of a CD47xCD19 κλ body was evaluated in a Rajimodel of lymphoma. 2.10⁶ Raji cells were implanted subcutaneously inNOD/SCID mice. Tumor volumes were measured 3 times per week. After thetumor graft reached 0.1 cm³, mice were randomized into 5 groups (5 miceper group) and the antibody treatment was initiated. This experimentcompared the effect of CD47xCD19 κλ-body Ka3xD11 to the effect of Ka3monovalent antibody, and two positive control Mabs, the CD47 Mab B6H12and rituximab. Antibody was injected i.p. three times per week until theend of the experiment (d25). Rituximab was administered at 200 μg permouse per injection. All the other antibodies were administered at 400 gper mouse per injection.

As shown in FIG. 13 the efficacy of the CD47xCD19 κλ-body Ka3xD11 issimilar to B6H12 known to bind strongly to CD47, block CD47-SIRPαinteraction and to suppress tumor growth in this lymphoma model. Ofnote, the efficacy of the CD47xCD19 κλ-body was also comparable to theefficacy of rituximab. The monovalent CD47 antibody was clearly lessefficacious than the CD47xCD19 bispecific κλ-body demonstrating thattumor eradication is CD19-dependent.

Example 15: CD47 Antibody Binding to Erythrocytes

With more than 5 billion cells per ml of blood, and 25,000 CD47molecules per cell, erythrocytes represent potentially the major antigensink for CD47-binding antibodies. To assess the effect of erythrocyteadsorption, CD47 antibodies were incubated with whole blood. Followingincubation, the fraction of CD47 antibodies remaining in the plasma wasdetermined by ELISA.

In brief, 200 μl of whole blood containing an anti-coagulant was mixedwith 20 μl of antibody (110 μl/ml in PBS) and incubated for 30 minutesat 37° C. with shaking. The plasma was then separated from the cells bycentrifugation, and the concentration of unbound antibody determined byELISA. For each antibody tested, the results obtained were compared tothe control, that is the same antibody spiked directly into plasma, andnormalized against non-binding IgGs tested in parallel.

FIG. 14 demonstrates that high and moderate affinity CD47 antibodies areefficiently adsorbed on erythrocytes. However, in the case of BsAbs,this phenomenon is limited to molecules having a high affinity CD47arms, such as 5A3. This suggests that, in general, BsAbs are less proneto erythrocyte adsorption and TMDD than CD47 Mabs.

OTHER EMBODIMENTS

While the invention has been described in conjunction with the detaileddescription thereof, the foregoing description is intended to illustrateand not limit the scope of the invention, which is defined by the scopeof the appended claims. Other aspects, advantages, and modifications arewithin the scope of the following claims.

What is claimed is:
 1. A bispecific antibody, comprising: a. a firstantigen binding arm that specifically binds CD47 and comprises: i. aheavy chain variable region comprising: a variable heavy chaincomplementarity determining region 1 (CDRH1) amino acid sequence of SEQID NO: 225, a variable heavy chain complementarity determining region 2(CDRH2) amino acid sequence of SEQ ID NO: 226, a variable heavy chaincomplementarity determining region 3 (CDRH3) amino acid sequence of SEQID NO: 227; and ii. a light chain variable region comprising: a variablelight chain complementarity determining region 1 (CDRL1) amino acidsequence of SEQ ID NO: 239, a variable light chain complementaritydetermining region 2 (CDRL2) amino acid sequence of SEQ ID NO: 242, anda variable light chain complementarity determining region 3 (CDRL3)amino acid sequence of SEQ ID NO: 254 or SEQ ID NO: 246; and b. a secondbinding arm that specifically binds CEA.
 2. The bispecific antibody ofclaim 1, wherein the antibody comprises a variable heavy chain aminoacid sequence of SEQ ID NO: 114 and a variable light chain amino acidsequence selected from SEQ ID NO: 168 or SEQ ID NO:120.
 3. Thebispecific antibody of claim 1, wherein the antibody comprises twocopies of a single heavy chain polypeptide and a first light chain and asecond light chain, wherein the first and second light chains aredifferent.
 4. The bispecific antibody of claim 3, wherein at least aportion of the first light chain is of the Kappa type and at least aportion of the second light chain is of the Lambda type.
 5. Thebispecific antibody of claim 4, wherein the second light chain comprisesat least a Lambda constant region.
 6. The bispecific antibody of claim5, wherein the second light chain further comprises a Lambda variableregion.
 7. The bispecific monoclonal antibody of claim 5, wherein thesecond light chain further comprises a Kappa variable region.
 8. Thebispecific antibody of claim 1, wherein the antibody is fully human. 9.The bispecific antibody of claim 3, wherein the first light chaincomprises a Kappa constant region and a Kappa variable region, andwherein the second light chain comprises a Lambda constant region and aLambda variable region.
 10. A method of treating, alleviating a symptom,preventing, or delaying the progression of pathologies associated withaberrant CD47 and/or aberrant CD47-SIRPα expression and/or activitycomprising administering to a subject in need thereof a compositioncomprising the bispecific antibody of claim
 1. 11. The method of claim10, wherein pathologies associated with aberrant CD47 and/or aberrantCD47-SIRPα expression and/or activity is cancer.
 12. The method of claim11, wherein the cancer is leukemia, lymphoma, breast cancer, coloncancer, ovarian cancer, bladder cancer, prostate cancer, glioma, lungcancer, bronchial cancer, colorectal cancer, pancreatic cancer,esophageal cancer, liver cancer, urinary cancer, bladder cancer, renalcancer, kidney cancer, pelvis cancer, oral cavity cancer, pharynxcancer, uterine corpus cancer or melanoma.